developing a porcine slaughterhouse model for normothermic...

Developing a porcine slaughterhouse model for normothermic regional perfusion of donor kidneys Student Vera Tichelaar Faculty supervisor prof dr HGD Leuvenink Daily supervisor LH Venema University Medical Center Groningen Department of Surgery ndash Surgical Research Laboratory 01-09-2016

A porcine slaughterhouse model for normothermic regional perfusion of kidneys

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Abstractsamenvatting Objectives To decrease waiting time for a kidney transplant donor kidneys of inferior quality

are increasingly being accepted Normothermic regional perfusion (NRP) in donation after

circulatory death restores abdominal circulation with autologous blood and improves organ

quality The use of blood in machine perfusion can cause inflammation injury tissues damage

and cell death Probably it is better to replace the blood in NRP with an artificial preservation

solution To test the different solutions a normothermic machine perfusion model for an

isolated kidney has to be designed The aim of this study is to design a NMP model with

porcine slaughterhouse kidneys to test kidney viability

Methods Porcine kidneys and autologous whole blood were obtained from the

slaughterhouse Kidneys were transported to our lab and reperfused for 4 hours using the

NMP set-up Seven groups were created (n=1-4) Kidneys were transported differently using

cold storage subnormothermic machineperfusion oxygenated- or non-oxygenated

hypothermic machine perfusion Warm and cold ischeamic times differ between groups In

one group mannitol insulin nutrients and dexamethson was added during NMP (NMP+)

Hemodynamics were monitored and perfusate and urine samples were taken regularly

Biopsies were taken to asses renal histology

Results A stable NMP was established There was a significant difference between groups

Kidneys preserved using oxygenated hypothermic machine perfusion and reperfused with

NMP+ performed significantly better These kidneys showed better creatinine clearance

sodium reabsorption and urine production than the control group

Conclusion The model created using oxygenated hypothermic machine perfusion and NMP+

likely is useful for testing different perfusion solutions However more research is required to

optimize this model

Doelstellingen Om wachttijd voor een niertransplantatie te verkorten worden donornieren

van mindere kwaliteit in toenemende mate geaccepteerd Normotherme regionale perfusie

(NRP) na donatie na hartdood hersteld de abdominale circulatie met autoloog bloed en

verbeterd de orgaan kwaliteit Het gebruik van bloed in machine perfusie kan inflammatoire

schade weefselschade en celdood veroorzaken Waarschijnlijk is het beter om bloed

tevervangen door een artificieumlle preservatie vloeistof Om verschillende vloeistoffen te

kunnen testen moet er een normotherm machineperfusie (NMP) model ontworpen worden

Het doel van deze studie is het ontwerpen van een NMP model met varkens nieren uit het

slachthuis om de kwaliteit van de nieren te testen

Methode Varkens nieren en autoloog bloed werden verzameld in het slachthuis De nieren

werden vervoerd naar ons lab en 4 uur op de NMP opstelling geperfundeerd 7 groepen zijn

getest met 1-4 nieren per groep Er zijn verschillende manieren van transport gebruikt cold

storage subnormotherme machineperfusie geoxygeneerde of niet ge-oxygeneerde

hypotherme machineperfusie Warme en koude ischemie tijden verschillen per groep In een

groep is tijdens NMP mannitol insuline voedingsstoffen en dexamethason toegevoegd

(NMP+) De hemodynamica werd gemonitord en perfusaat en urine monsters werden

regelmatig genomen Biopsieeumln werden genomen om renale histologie te beoordelen

Resultaten Een stabiel NMP systeem werd verkregen De groep waarbij geoxygeneerde

hypotherme machineperfusie en NMP+ gebruikt werd presteerde significant beter dan de

controle groep Er was betere creatinine klaring natrium reabsorptie en urine productie

Conclusie Het gecreeumlerde model waarbij gebruik gemaakt wordt van geoxygeneerde

hypotherme machineperfusie en NMP+ is bruibaar om verschillende vloeistoffen voor

perfusiedoeleinden te testen Verder onderzoek is nodig om de nierfunctie tijden de NMP

periode te verbeteren


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Table of contents Abstractsamenvatting 2

Introduction 4

Organ shortage 4

Normothermic regional perfusion 5

Perfusion solutions in NRP 6

Machine perfusion 7

Study objectives 8

Material and methods 9

Experimental design 9

Organ and blood retrieval 9

Kidney transport 10

Perfusion 11

Urine and perfusate analysis 13

Statistical analysis 13

Results 14

Stabilizing the NMP system 14

Renal hemodynamics 16

Renal function 18

Renal Histology 22

Discussion 23

Considerations 23

Study strengths and limitations 24

Recommendations for future research 25

Bibliography 26

Acknowledgements 29

Appendices 30


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Introduction

Organ shortage Kidneys are the most frequently transplanted solid organs Patients suffering from end-stage

renal disease kidney transplantation offers improved quality of life and better life expectancy

when compared to dialysis The persisting organ shortage represents a severe problem in

transplant medicine1 (Table 1) Transplant waiting lists are increasing at a greater rate than the

availability of donors Declining numbers of the classical donation after brain death (DBD)

donors are available due improvements in the management of severe neurologic injuries2

This has triggered interest in marginal donors as an additional organ source Expanded criteria

donors (ECD) and donation after circulatory death (DCD) are examples of such marginal

donors Marginal donors that normally would have been declined in the past are now

considered for transplantation to decrease organ shortage3

Therefore the number of organ

donors has remained more of less stable However on the other hand a decreasing number of

patients become eligible for organ transplants due to obesity excessive alcohol consumption

poorly controlled hypertension and diabetes4 Thereby maintaining the long waiting time for a

kidney transplant Kidney transplantation outcome is negatively affected by this waiting time

with poorer outcome for patients subjected to prolonged dialysis5

A possible solution for organ shortage is the use of donation after circulatory death donors

The different types of DCD may be categorized using the Maastricht criteria6 (Table 2) The

controlled DCD- or Maastricht category III donors are most used in the Netherlands and

are those who have suffered massive brain injury but do not meet the criteria of brain death A

decision to withdraw supportive treatment is made independently of donor status Kidneys

from these patients undergo a period of warm ischemia between asystole and organ retrievel

leading to poorer transplant outcomes with higher incidence of primary non-function (PNF)

and increased complications rates7 In a porcine study an increasing warm ischemic time

(WIT) leads to a proportional impairment of early renal function associated with greater

severity of underlying oxidative tissue injury Kidneys sustaining less warm ischemia

demonstrated better function represented by creatinine clearance urine output renal

hemodynamics and oxygen consumption8

Table 1 Kidney graft shortage in US and Eurotransplant region

United states

(National Kidney

Foundation)

Eurotransplant region

(Eurotransplant

international foundation

statistics report)

Patients on kidney

transplant waiting list

100791 (January 2016) 10282 (March 2016)

Deceased donor kidneys

transplanted

11570 (2014) 1827 (2015)

Median waiting time to

deceased donor kidney

transplant

Up to 5 years Up to 4 years


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Table 2 Maastricht Categories of Donation after Circulatory Death

Category Description

I Dead on arrival at the hospital

II Unsuccessful resuscitation at the hospital

III Withdrawal of supportive treatment

IV Cardiac arrest following establishment of brain death

Patients who have circulatory arrest in relatively uncontrolled situations may also become

cardiac death donors These ldquouncontrolled DCD-rdquo or Maastricht categories I en II donors

experience a longer period of warm ischemia than controlled DCD which results in even

higher incidences of PNF and delayed graft function (DGF)7

Normothermic regional perfusion Organ procurement from DCD donors is associated with a higher rate of organ injury and

discards most likely due to the haste of removing the organs to minimize the WIT9

Normothermic regional perfusion (NRP) is a new and advanced technique which can be

utilized in DCD donors It restores the abdominal circulation with oxygenated blood in situ

between asystole and procurement This permits dissection without ischemic injury since

oxygen supply to the abdominal organs is guaranteed It also allows assessment of the organs

run tests introduce therapies if necessary NRP relies on an adequate supply of oxygen and

other substrates to fuel processes of cellular homeostasis as well as repair Given that cellular

metabolism is fully restored normothermic perfusion allows a more comprehensive

assessment of organ viability prior to recovery and transplantation1011

The use of NRP to facilitate organ donation was first described in 199712

NRP has been

developed in Spain for uncontrolled DCD donors (Maastricht II DCD II) where it increased

the donor pool The reported experience indicates low rates of PNF and a reduction in DGF

with good 1-year graft survival in kidney transplantation1314

NRP is likely to reduce the rate

of damage caused by warm ischemia as it re-establishes the abdominal circulation allowing a

careful identification of the vascular structures and enables the procedure to be performed

without undue speed compared to traditional DCD organ recovery15

During warm ischemia

ATP degradation leads to the progressive accumulation of xanthine and hypoxanthine

important sources of superoxide radicals at organ reperfusion A period of NRP after warm

ischemia helps to restore cellular energy substrates reduce levels of nucleotide degradation

products and improve the concentrations of endogenous antioxidants16

Maintaining

circulation before retrieval is also thought to condition the organs with the up-regulation of

adenosine receptors which may protect against preservation injury17

The use of NRP in DCD II donors is associated with a lower risk of DGF and with a better

graft function 2 years post-transplantation compared to expanded criteria donor (ECD)

kidneys ECD are defined as donors over 60 years old or aged 50-59 years with at least two

of the following conditions cerebrovascular cause of death serum creatinine over 15 mgdL

or hypertension18

Organs from ECD were associated with a suboptimal post-transplantation

function or shorter graft survival A study by Demiselle et al compared patient survival graft

survival and kidney function between DCD II without NRP ECD and standard criteria

donors The post-transplantation results of DCD II kidneys were comparable to those of ECD

kidneys NRP preservation may improve the results of DCD II transplantation19

Furthermore

the feasibility of NRP in category III DCD donation has also been tested and it is possible to

establish NRP successfully and continue normothermic perfusion for a period of 2 hours In


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situ NRP represents a significant advance in DCD organ retrieval and has the potential to

increase the number and quality of the transplanted organs15

Perfusion solutions in NRP Different techniques have been developed to accommodate NRP in DCD II and DCD III

donors The use of NRP in both donation techniques requires a perfusion fluid which

comprises all the components needed during the NRP period The composition of the

perfusion solution is vital to ensure adequate delivery of nutrients and oxygen to maintain

cellular integrity and vascular processes

To ensure that sufficient oxygen for normal metabolic function is provided to the abdominal

organs an oxygen carrier in the perfusion solution is needed All normothermic perfusion

(NP) studies used blood as the perfusion solution with red blood cells as the oxygen

carrier158 NP with oxygenated blood was able to restore depleted ATP levels and reverse

some of the deleterious effects of CS20

Although red blood cells are highly evolved to provide oxygen to tissues there are some

disadvantages to using blood as perfusion fluid Early studies found that leucocytes

haemolysis and platelet activation during perfusion with a blood-based solution caused an

increase in resistance and tissue oedema during prolonged periods of preservation20

Leucocytes play a role in an inflammatory process causing cellular injury Endothelial cell

damage caused by ischemic injury stimulates a pro-inflammatory environment which

activates and stimulates leucocytes Leucocytes migrate and infiltrate into the interstitium

leading to microvascular congestion and the ldquono-reflowrdquo phenomena This increases cytokine

expression production of oxygen free radicals and activates the complement system to sustain

the injury response causing cell death and tissue damage21

Normothermic perfusion using a leukocyte and platelet depleted red cell-based solution limits

infiltration and the inflammatory response to improve circulation and renal function

Apoptosis and inflammatory mediators are also suppressed reducing the likelihood of

injury22

Platelets also have a damaging role in reperfusion injury they mediate

vasoconstriction and inflammatory processes causing injury23

In cardiovasculair surgery the use of cardiopumonary bypass (CBP) is associated with acute

kidney injury (AKI) Haemolysis is a common consequence of cardiopulmonary bypass that is

caused by mechanical stress in the perfusion circuit and results in the release of hemoglobin

from lysed erythrocytes into the plasma Cell-free oxyhemoglobin reacts with nitric oxide to

form methemoglobin and nitrate As nitric oxide is an important vasodilator that has a central

role in blood flow regulation reduction of nitric oxide bioavailability by free hemoglobin may

impair tissue perfusion24

Since the NRP circuit consists of the same technical features as a

cardiopulmonary bypass circuit this problem with AKI seen in CPB could be a serious

detrimental effect that can be the result when using NRP in combination with blood

Currently blood is still the widely used product in perfusion solutions for NRP and other

normothermic machine perfusion (NMP) settings As described above the components of a

blood-based solution causes inflammatory injury tissue damage and cell death Also

mechanical stress in a perfusion circuit contributes to kidney damage Therefore we believe

that the use of blood in machine preservation of organs is not the best solution and that is

would be better to replace the blood with another preservation solution

The artificial perfusion solution has to meet some requirements The perfusion fluid should

deliver enough oxygen to maintain aerobic metabolism Furthermore it needs to consist


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sufficient nutrients to prevent depletion of cellular energy substrates With these components

the preservation fluid should minimize injury in organs that have been subjected to warm

ischemia

In an ideal situation the non-blood based NRP solutions are tested in a randomized controlled

trail with NRP in DCD donors However it is unethical to do this in a clinical setting

Therefore the first step is to design an animal model Porcine kidneys are suitable for this

model because the size and geometry of a porcine kidney is comparable to human kidneys

Furthermore various renal functions damage parameters and morphology can easily be

assessed25

Performing NRP in a pig is an expensive procedure large experimental animals

are costly to keep and NRP equipment is expensive as well Also approval for such

experiments is hard to get in the Netherlands Therefore we are aiming to establish a low cost

model using slaughterhouse kidneys instead of laboratory pigs thereby decreasing the cost

and avoiding ethical questions However it is not possible to perform NRP in the

slaughterhouse therefore slaughterhouse kidneys are transported to our lab and tested in and

an isolated perfused porcine kidney system created to simulate warm kidney perfusion

Machine perfusion Preserving the function of the kidney graft during transport is of vital importance for an

effective NRP model In the clinical setting in most countries a kidney is flushed cooled with

a cold preservation solution and cold stored on ice between organ retrieval and

transplantation26

During this preservation period the organ is transported cross matching is

performed and the operating room can be prepared This period of cold ischemia is then

followed by reperfusion More and more research is performed to determine the best

preservation method between organ retrieval and transplantation The goal in these studies is

to decrease the amount of ischemia-reperfusion injury (IRI) caused by tissue ischemia27

IRI is an unavoidable relevant consequence after kidney transplantation and results in a

distinct inflammatory reaction of the graft Clinically IRI is associated with delayed graft

function graft rejection chronic rejection and chronic graft dysfunction28

IRI is principally

caused by blood flow impairment which starts with brain death and is due to severe

hemodynamic disturbances in cadaveric donors Clamping of the renal artery during the

harvesting operation causes a short but severe renal ischemia In addition cold ischemia

during transport causes a further ischemic damage The final and biologically more severe

stage of the injury occurs during the reperfusion as a consequence of the returning blood flow

in the recipient29

Underlying factors of ischemia reperfusion include energy metabolism

cellular changes of the mitochondria and cellular membranes initiation of different forms of

cell death-like apoptosis and necrosis together with a recently discovered mixed form termed

necroptosis Chemokines and cytokines together with other factors promote the inflammatory

response leading to activation of the innate immune system as well as the adaptive immune

system If the inflammatory reaction continues within the graft tissue a progressive interstitial

fibrosis develops that impacts long-term graft outcome30

Machine perfusion could enable active organ conditioning prior to transplantation and

furthermore this technique provides a platform for therapeutic interventions during organ

preservation21

Machine perfusion is preferably done under (sub)physiologic conditions

through (sub)normothermic machine perfusion at or below 37degC31

NMP may be able to

reverse some effects of ischemia by restoring organ metabolism outside the body prior to

transplantation It also allows pre-transplant assessment of organ viability25

Kidneys that


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have been perfused using NMP have significantly lower rates of DGF than those preserved

cold storage33

Various studies have demonstrated that hypothermic machine perfusion (HMP) is superior to

static cold stored kidney grafts from deceased donors 2634

DCD donors are more likely to

suffer from IRI only cooling the organ during preservation may not be sufficient The

principle of cold preservation is based on temperature reduction to reduce metabolism

Cooling does not completely stop cell metabolism which in turn leads to energy depletion35

HMP however reduces the risk and duration of DGF and leads to improved graft survival26

In response to these convincing data all kidneys recovered from deceased donor kidneys in

The Netherlands are preserved by HMP as of November 2015 Static cold storage has been

largely abandoned in our country for kidney preservation36

However the need for oxygen during HMP persists because the metabolic rate remains at

levels estimated around 10 There has been much debate on whether it is necessary to add

oxygen to support the low level of metabolism under these conditions Evidence suggests that

oxygen is particularly beneficial in restoring cellular levels of adenosine triphosphate after

kidneys have been subjected to warm or cold ischemic injury37

The potential benefits of

active oxygenation during HMP have been tested using a pig model Oxygen delivery during

preservation proved to be valuable for improving organ quality Kidney grafts preserved with

oxygenated HMP displayed a lower serum creatinine peak compared to non-oxygenated

HMP Histologic investigation showed a trend towards decreased inflammation in kidneys

preserved with oxygen38

Study objectives The aim of this study is to design a NMP model with porcine slaughterhouse kidneys to test

kidney viability The results of this study serve as a basis for the development of a preclinical

study where different perfusion solutions for NRP will be tested and later verified in a large

animal model

The first priority is to establish a stable perfusion using the IPPK technique for 4 hours The

NMP system should be pressure controlled and maintain a mean pulsatile arterial pressure of

75 mmHg The perfusate in the system must be 37 degC to represent normal physiological body

temperature The oxygenator should be able to deliver enough oxygen to the perfusate to keep

the partial oxygen pressure above 60 kPa

Furthermore the optimal way to preserve the porcine kidneys from the slaughterhouse to the

lab needs to be explored First different WIT will be tested Secondly a different method of

transportation will likely improve kidney quality This is tested by using cold storage

subnormothermic oxygenated machine perfusion hypothermic non-oxygenated machine

perfusion hypothermic oxygenated machine perfusion In the end we will add additives

during reperfusion to support kidney function


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Material and methods

Experimental design

Eight groups were created with 2 kidneys each except for the control group which contains 4

kidneys The WIT way of transport and cold ischemic time (CIT) differs between the groups

as described in table 3 Kidneys were transported differently either using cold storage (CS)

subnormothermic machineperfusion (sNMP) oxygenated hypothermic machine perfusion for

3 hours or non-oxygenated hypothermic machine perfusion for 3 hours(HMP -O2) All

kidneys were reperfused for 4 hours in a normothermic machine perfusion (NMP) set-up at an

arterial pressure of 75 mmHg and temperature of 37degC In the last group the NMP protocol

has changed dexamethson and mannitol was added to the priming solution and insulin

nutrients and bicarbonate were added during perfusion to create a NMP+ group Renal blood

flow and perfusate temperature were recorded every 10 minutes Perfusate and urine samples

were taken every 30 minutes Both were kept on ice before centrifugation and storage at -80C

Blood gas samples were taken and analysed immediately every 30 minutes One needle

biopsy of the cortex was taken prior to perfusion and a surgical biopsy was taken after

perfusion

No cold ischemia in this group because preservation of the kidney was with oxygenated

subnormothermic machine perfusion

Organ and blood retrieval Kidneys were retrieved from two different slaughterhouses in the vicinity of Groningen The

protocol for organ and blood retrieval was the same for both (appendix 1) Pigs were

anaesthetized with a bi-temporal electric shock rapidly followed by exsanguination

following standard slaughterhouse procedures under the supervision of a veterinarian

Approximately 3 liters of autologous blood was collected in a beaker containing 5ml25000

units of heparin The blood was then poured into a jerry can for transport The kidneys were

removed and after a warm ischeamia interval one was flushed with NaCl 09 until the

aspect of the kidney became uniformly pale and clear fluid ran from the vein The kidney was

then stored for transport according to the assigned group Cold ischemic times varied with the

experimental groups

Table 3 experimental groups

Group N= WIT (min) Transport CIT(hour) Reperfusion

1 (control) 4 30 CS 3 NMP

2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +


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Kidney transport After removal and flush of the kidneys they were stored differently using CS HMP or sNMP

For CS after flushing with cold NaCL 09 the kidney was stored in an organ bag containing

NaCL 09 and stored When HMP was applied the kidney was flushed with cold NaCL

09 and connected to a hypothermic machine perfusion pump (Kidney Assist transport

Organ Assist Groningen The Netherlands) seen in figure 3 filled with cold UW-MP

solution (belzers MP Bridge to life Londen United Kingdom) A patch was created using the

aorta and placed in a patch holder and connected to the kidney holder This is shown in figure

1 figure 2 and figure 3 After this the kidney was placed on the machine Hypothermia was

maintained because of crushed ice surrounding the circuit in which the kidney is placed The

oxygen bottle of the device was opened according to the experimental group

Figure 1 Kidney with patch Figure 2 Patch connected to patch holder

Figure 3 Placement in kidney holder


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For sNMP the kidney was also connected to a Kidney Assist transport (KA) (Organ Asisst

Groningen Netherlands) only instead of ice surrounding the circuit the machine is filled with

heat packs and primed with 500 ml autologous whole blood and 500 ml ringerslactate and

perfused at a temperature of 30degC After flushing with warm NaCL 09 excess fat was

removed and the ureter vein and artery were cannulated then placed in a kidney holder and

placed in the KA reservoir

Figure 4 Kidney assist with disposable

Attaining leukocyte depleted autologous whole blood The leukocyte-depleted blood was prepared by filtering the heparinised autologous whole

blood collected at the slaughterhouse First the blood was poured in a catheter bag using a

funnel Then the blood was led through a leukocyte filter (BioR O2 plus Fresenius Kabi

Zeist Netherlands) After filtration the blood was checked for leukocytes with an upper

boundary off 001x10^9

Perfusion The perfusion circuit that was designed contains a KA with a centrifugal pump (Medos

Medizintechnik AG Stolberg Germany) an oxygenator (Hilite 800 LT Medos

Medizintechnik AG Stolberg Germany) and a homemade organ chamber with a cannula

(cannula for organ perfusion ndash 12F INFUSION Warszawa) To keep the perfusate

temperature stable at 37degC an oxygenator with integrated heat exchanger was used A

temperature sensor provided information regarding the temperature Flow was monitored

using an ultrasonic clamp-on flow probe (ME7PXL clamp frac14 inch flow meter Transonic

Systems Inc Ithaca NY) Pressure was measured directly before the cannula using pressure

transducer which was zero-calibrated to the atmosphere (TrueWave disposable pressure

transducer Edwards Lifesciences Irvine CA) All components were attached to each other

using disposable tubing (Rehau Rauclair-E 102 10x14 and 715 7x10 Rehau NV Nijkerk

Netherlands) (appendix 2) The circuit is shown in figure 5


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Figure 5 The perfusion circuit

The set up was primed with 300 ml Ringers Lactate 10 ml voluven 10ml bicarbonate 100microl

sodium nitroprusside (20mgml) and amoxicillin-clavulanate 1000mg200mg (Sandoz BV

Almere Netherlands) Creatinine was added to achieve a concentration of 1 moll After

priming 500 ml leukocyte depleted whole blood was added The perfusate was oxygenated

with 05 Lmin carbogen (95 O2 5 CO2)

Preparation of the kidney was initiated when the perfusate was 37degC Excess fat was removed

and the ureter was cannulated with an 8 Fr nasogastric feeding tube (Nutrisafe 2 gastro-

duodenal feeding tube (Pur) 8Fr Vygon Valkenswaard Netherlands) The artery patch was

removed and the artery was cannulated with an arterial cannula The cannulated kidney is

shown in figure 6 Next the kidney was put in the organ chamber and attached to the

perfusion circuit (figure 7) and perfused in a pulsatile sinusoid fashion at a mean arterial

pressure of 75 mmHg for a total duration of 4 hours

Figure 6 Cannulated Kidney Figure 7 Kidney connected to NMP circuit


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To create the 30WI+HMPO2+NMP+ group two infusion pumps are added to the circuit

These pumps are connected to the oxygenator using a valve system For this group the

priming solution is altered 6mg mannitol and 6 mg dexamethason is added The infusion

pumps are used to infuse a nutrient solution with added insulin at 20mlhour and glucose

09 at a rate of 7 mlhour

Histology All pre- and postperfusion biopsie were fixed in 4 formalin dehydrated and embedded in

paraffin wax Sections were cut then stained with hematoxylin and eosin (HE) for evaluation

using light microscopy

Urine and perfusate analysis Urine and perfusate were analysed with routine automated test methodology carried out by the

clinical diagnostics laboratory after completing all experiments Creatinine and sodium levels

were determined in every sample both in urine and perfusate Creatinine clearance (=(urine

creatinine concentration x urine flow rate) plasma creatinine concentration) and fractional

excretion (=100 x (Sodium urine concentration x plasma creatinine concentration) (plasma

sodium concentration x urine creatinine concentration)) of sodium were calculated Lactate

dehydrogenase (LDH) was also determined in a number of experiments as marker of

generalized cellular stress (Table 4)

Statistical analysis Values are presented as mean with standard deviations Descriptive statistics were used to

display statistical dispersion of kidney function parameters within each group Continuous

variables such as serum creatinine were plotted as level versus time curves for each kidney

and the mean area under the curve (AUC) was calculated An one-way ANOVA was used to

compare values between groups if the data were normally distributed and had homogeneity of

variances If data failed these assumptions the Kruskal-Wallis H test was used P-values le

005 were assumed to indicate statistical significance Post hoc tests were performed if

necessary

Table 4 Viability assesment

Perfusion

parameters

Renal function Tubulair function Injury markers

Perfusion pressure Serum creatinine

levels

GFR LDH

Flow Creatinine clearance Fractional NA

excretion

Lactate

Oxygen concetration pH

Kidney weight ATP


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Results

Stabilizing the NMP system The first 4 kidneys that were perfused were used to stabilize the NMP system to our

requirements The results were analysed after perfusion and adjustments were made to the

system or perfusate when necessary The Kidney Assist was able to provide a stable 4 hour

pressure controlled perfusion at 75 mmHg The third kidney was excluded from the analysis

The decision was made to stop the experiment when the oxygenator started to leak vigorously

Perfusate temperatures renal blood flow and diuresis are shown in the table below

The water bath and heat chamber were able to warm-up the perfusate temperature to 37degC

When connecting a cold stored kidney to the perfusion circuit a temperature drop is seen after

which the temperature is increasing to the appropriate level To maintain stable temperatures

sample were taken via a hatch in the surrounding cabinet instead of taking the entire front of

The blood flow values were low in the first two experiments Therefore a vasodilator was

added to the priming solution This resulted in higher blood flows and more diuresis in the

forth experiment (table 6) which was more in line with expectations for a porcine kidney

Table 5 Perfusate Temperature (degC) Renal bloodflow (mlmin) and diuresis (ml) of first 2 kidneys

Kidney 1 Kidney 2

Time Temperature Flow Diuresis Temperature Flow Diuresis

t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62

Table 6 Perfusate Temperature (degC) Renal bloodflow (mlmin) and diuresis (ml) of kidney 4

Time Temperature Flow Diuresis

t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


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After analysing the perfusate a number of improvements were made to create an environment

for the kidneys that was as close to physiological as possible First the partial oxygen pressure

was checked using gas analysis Graph 1 shows that the oxygenator can keep the oxygen level

above 60 kPa

Graph 1 Oxygen pressure in Perfusate

Glucose levels were also monitored in the perfusate during perfusion (graph 2) During the

first experiment glucose levels dropped until 02 mmolL This level is insufficient to support

normal cell metabolism Therefore we added 7 ml 09 glucose hourly in the second

experiment The goal was to achieve a concentration of 8 mmolL The glucose levels during

the second were higher but did not reach the 8 mmolL goal In the fourth experiment we

calculated the amount of glucose 09 needed to be added to increase the concentration up to

8 mmolL at each time point which gave better results In the following experiments we used

the same table and added glucose 09 before starting perfusion to increase the glucose level

at t=0

Graph 2 Glucose concentration in Perfusate

40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


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16

Renal hemodynamics Kidneys 5 to 22 were used to fill the experimental groups The control group and HMP+O2

group consisted of 4 kidneys the other groups had 2 kidneys each Except for the

30WI+HMP+ O2+NMP+ which has only one kidney The second kidney in this group was

excluded from analysis due to a broken pressure sensor We could not start HMP preservation

until replacing the sensor During the time it took to replace the pressure sensor the kidney

was cold stored instead of HMP preserved When reperfused renal blood flow of this kidney

was much lower than other kidneys which also led to poor ability to re-warm the kidney to

37degC Not meeting the standards set for the experimental group led to exclusion of this

kidney

Renal blood flow increased during the first 30 minutes in all groups After this the flow

remained almost constant until the last two hours in which the flow is gradually decreasing

Mean flow per group with standard deviation is presented in graph 3-9 Each time point is

evaluated using a one-way ANOVA there were no significant differences found P-values of

the statistical analyses are shown in table 7

Graph 3-6 Mean Renal blood flow in mlmin100 gram per experimental group


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17


Table 7 statistical analysis of renal blood flow per time point using one-way ANOVA

Time point (min) Significance difference (p-value)


0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


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18

Renal function Urine production was collected every half hour Graph 4-10 shows mean urine production and

standard deviation per experimental group The high urine production of the

30WI+HMP+O2+NMP+ group suggests a better performance of this kidney Statistical

analysis of all groups using a Kruskal-Wallis H test showed that at t=120 and t=150 a

significant difference is present (table 8) Therefore a post hoc test is performed for both time

points The 30WI+HMP+O2+NMP+ has significantly more urine production compared to the

control group at t=120 and t=150 p=0001 and p=0002

Graph 10 Mean urine production in mlmin per experimental group

Table 8 statistical analysis of urine production per time point in all groups using a Kruskal-Wallis H test


15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


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19

As a mark for kidney function creatinine clearance and the fractional sodium excretion

(FENa+) were calculated using creatinine and sodium levels in perfusate and urine Mean

creatinine clearance per group is presented in graph 11 Creatinine clearance rates were

analysed using either a Kruskal-Wallis H test or a one-way ANOVA results are shown in

table 9

Graph 11 Mean creatinine clearance per experimental group

At t=15 t=90 t=120 t=180 and t=210 significant results appeared which needed further

evaluating The 30WI+HMP+O2 and 30WI+HMP+O2+NMP+ groups have a significantly

better creatinine clearance level compared to the control group When comparing these last 2

groups there is a significant difference at t=15 and t=90 indicating the

30WI+HMP+O2+NMP+ is even better than the 30WI+HMP+O2 group Post hoc results are

presented in table 10

0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)

Creatinine clearance

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

Table 9 statistical analysis of creatinine clearance per time point

Time point (min) Significance difference (p-value) Test used

15 0000 One-way ANOVA

30 0252 Kruskal-Wallis H









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Table 10 Post Hoc analysis of creatinine clearance per time point

Timepoint (min) 30WI+CS vs 30WI+HMP+O2

30WI+CS vs 30WI+HMPO2+NMP+

30WI+HMP+O2 vs 30WI+HMPO2+NMP+

15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165

The serum creatinine drop after 4 hour NMP was calculated for each group The

30WI+HMP+O2 and 30WI+HMP+O2+NMP+ group cleared a significantly better percentage

of creatinine then our control group (p=0007 and p=0001) There was no difference when

comparing the 30WI+HMP+O2 with 30WI+HMP+O2+NMP+ (p=0436)

All mean FENa+ are plotted in graph 12 The FENa

+ of the 30WI+CS and 20WI+CS were

high suggesting that kidney function is less than other groups The 40WI+sNMP group

appears to be better than the other groups during the first hour however this can be explained

by the fact that one kidney in this group did not produce any urine for the first hour

Graph 12 Mean fractional excretion of sodium per experimental group

When evaluating these values using a Kruskal-Wallis H test or an one-way ANOVA results

show a significant difference at all time points after t=90 (table 11) Post Hoc testing reveals

that after t=90 the 30WI+HMP+O2 and 30WI+HMP+O2+NMP+ are functioning significantly

better then the control group However comparing the 30WI+HMP+O2 group with the

30WI+HMP+O2+NMP+ group there is no significant difference present as seen in table 12

Since the groups were small the area under the curve (AUC) for FENa+ was calculated and

analysed using an one-way ANOVA This showed a significant difference p=0027 Post hoc

analysis revealed both the 30WI+HMP+O2 and 30WI+HMPO2+NMP+ group were

0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)

Fractional exrection of sodium

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

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21

significantly better than the control group There was no significant difference between those

two groups

Several other kidney function and tissue injury parameters were evaluated and an overview is

presented in appendix 3 Kidneys were weighed before and after NMP Weight gain is the

highest in the 40WI+sNMP group Lactate and LDH are also analyzed Lactate levels are

increasing during the 4 hours reperfusion except in the 30WI+HMPO2 group were lactate

levels are decreasing Other parameters such as pH pO2 and glucose did not differ between

groups and are also presented in appendix 3 There are no more statistically significant

differences than previously discussed in kidney function and injury markers between groups

as shown in table 13

Table 11 statistical analysis of FeNa+ per time point










Table 12 Post Hoc analysis of FeNa+ per time point




90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548

Table 13 statistical analysis of kidney function and tissue injury parameters

Variable Significance difference (p-value)

Creatinine clearance (mlmin100gr) AUC 0054

Edema 0279

FeNa+ () AUC 0027

LDH concentration (UL) AUC 0648

Serum creatinine drop () 0001

Urine production accumulated (ml) 0076


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22

Renal Histology Apart from analyzing renal hemodynamics and kidney function parameters we also studied

renal histology of the biopsies A slight difference seemed to occur between groups A t=0

biopsy was taken after preservation before NMP All groups show damaged tubular cells

indicating that acute tubular necrosis (ATN) is present In the CS groups (figure 8) ATN is

more severe than the kidneys preserved with HMP (figure 9) Focal tubular epithelial necrosis

is present and rupture of basement membranes and occlusion of tubular lumens is more severe

in these groups

Figure 8 HE staining at t=0 of CS kidney Figure 9 HE staining at t=0 of HMP kidney

After 4 hours NMP a second biopsy was taken The difference between groups became more

evident evaluating the histology In the CS group most tubules were fully obstructed due to

necrosis of epithelial cells which have detached and sloughed into the tubular lumens Some

tubules appeared relatively normal meaning there was probably some function left Inside

Bowmanrsquos capsule protein deposition was present indicating that the glomeruli were leaking

There is no difference between the histology when WI changes The 40WI+sNMP group did

not differ from the CS group since debris and obstructed tubules are also present The HMP

groups showed open and intact tubules indicating better function as the CS and 40+sNMP

group The oxygenated kidneys had more arearsquos with almost normal tubules then the non-

oxygenated kidneys The best of all is the 30WI+HMPO2+NMP+ which showed more open

tubuli with a larger diameter and Bowmanrsquos space appeared better then all previous described

groups



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23

Discussion We showed that is it possible to develop a stable NMP system by which renal function

parameters can be monitored A pulsatile mean arterial pressure of 75 mmHg is maintained

during 4 hours perfusion at 37 degC The partial oxygen pressure stayed above 60 kPa This

NMP system is suitable to use as a porcine DCD model without using laboratory animals

Further experiments showed that kidneys in the 30WI+HMP+O2+NMP+ group had

significantly better results than our 30WI+CS control group

Considerations In this study we tried to create a NMP system that is stable and useful to test different

perfusion solutions later on When considering renal function a few parameters stood out

First of all we found a relatively low level of creatinine clearance during NMP Other studies

reported a much higher level of creatinine clearance up to 20 mlmin100gr394041

As we

know that warm and cold ischemia are detrimental to the kidney the short warm ischemia

time (6-7 minutes) and relatively short cold ischemia (2 hours) that the kidney were exposed

to in that study could provide a feasible explanation as to why there is such a large difference

in creatinine clearance However in our study we had a similar experimental group with 7

minutes warm ischemia and 2 hours cold storage Creatinine clearance in our group reached

only 5 mlmin100gr This difference could be due to variations in organ retrieval and

reperfusion protocols used in our experiments

Prolonged warm ischemia time is associated with graft failure and mortality after kidney

kidney transplantation7 Also a clear association between increasing warm ischemic time and

more severe IRI and deterioration in renal function has been shown8 However in our results

different WIT did not lead to statistical significant differences This is most likely caused by

the slaughter process The pigs experience a lot of stress during transport and also waiting for

their turn to be exsanguinated Another element is the heat drum used in standard

slaughterhouse procedures for removing hair and softening the skin adding extra warmth

during the ischemic period The small number of kidneys in each experimental group could

also have contributed to the statistical outcome more inclusions could strengthen statistical

tests and reveal significance if present

Also FENa+ values were investigated after NMP These values were spread between

extremely high and close to normal physiological levels The high FENa+ values are most

likely the result of ATN which is also seen in other studies3941

FENa+ is the highest in the

20WI+CS group (FeNA t240 = 8236plusmn471) and lowest in the 30WI+HMP+O2+NMP+ group

(FeNA t240 = 435) Comparing HE staining of both groups support this assumption In the

20WI+CS group there is more tubular damage evident than the 30WI+HMP+O2+NMP+

group

In addition all kidneys showed an increase in weight suggesting oedema formation This is

probably due to ischemia-reperfusion damage leading to intracellular and interstitial swelling

which is also seen in other studies3925

A point of interest from our view was whether adding oxygen to hypothermic machine

perfusion is beneficial during transport A study evaluating oxygenated hypothermic machine

perfusion in a DCD model showed that preservation using oxygenated hypothermic machine

perfusion is efficient in preserving DCD kidneys greatly enhancing the capacity of the graft

to withstand preservation stress and improving outcome38

Re-evaluating results from only the

30WI+HMP+O2 and 30WI+HMP-O2 group revealed a statistical difference in FENa+ after


Ho

ofd

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n

24

25 hour NMP (t=180 P=0025 t=210 p=0022 t=240 p=0042) However these values we

report are of very early renal function long term results could differ

Interesting are the results of the 30WI+HMPO2+NMP+ group To our belief this is the best

performing kidney Adding mannitol dexamethason to the perfusate and infusing nutrients

and insulin during perfusion is be beneficial to the kidney Mannitol has protective effects

including increasing renal blood flow and decreasing intravascular cellular swelling

Dexamethason is used to reduce the inflammatory response and insulin to stimulate the re-

absorption of glucose Apart from glucose as energy source it is likely that kidneys need

amino acids to build new proteins Other studies have better results using these additives

during experiments 323941

Our analysis shows a difference between the

30WI+HMPO2+NMP+ group and the control group We experienced some difficulties during

one of the experiments due to a broken pressure sensor Excluding this kidney still resulted in

a highly significant findings when evaluating statistics between the 30WI+HMP+O2 and

30WI+HMPO2+NMP+ group Indicating 30WI+HMPO2+NMP+ has better function

compared to 30WI+CS and 30WI+HMP+O2 However to properly evaluate the effects of

these additives more experiments and further investigation is necessary

Study strengths and limitations This study has several strengths First of all a major advantage was that this study is

performed using kidneys from commercial slaughterhouses making the use of laboratory

animals unnecessary Porcine kidneys resemble human kidney closely in function and

anatomy Normally a typical model utilizes laboratory animals as organ donors which is

associated with considerable costs Moreover sacrificing a laboratory pig to obtain one or two

kidneys for research may be regarded as inefficient and ethically questionable

We also succeeded in creating a stable model for testing possible improvements for DCD

donation with outstanding results in the 30WI+HMPO2+NMP+ group Although our results

are suboptimal compared to other studies this model is excellent for testing perfusion fluids

There is room for improvement in renal function which could be achieved by one of the

artificial perfusion solutions to be tested

There are also a few limitations of this study one of them being the small groups (n=1 n=2 or

n=4) The small number of kidneys per group makes is difficult to conclude what the effect of

different perfusion techniques are Due to little time and lots of different techniques to

evaluate we were not able to do more experiments per experimental group However most

interventions were also evaluated by other studies and we had to create similar results during

this pilot in order to show our experimental set up is functioning properly

During the last experiments we experienced some technical difficulties leading to delay

during set up or impairment during perfusion A broken pressure sensor caused delay in

machine preservation causing cold ischemia time instead of oxygenated machine perfusion

We also encountered some coagulation during some of the experiments The heater inside the

cabinet caused clotting in the ureter canula or line of the pressure sensor A blocked urethra

can cause congestion inside the kidney and impairment of kidney function When the pressure

sensor line is blocked pressure starts to build inside de pressure sensor and flow will be

regulated down unnecessary We also had some problems regarding oxygenation during some

experiments A leaky oxygenator made a oxygenator replacement required during or a few

minutes prior to reperfusion Fortunately once experienced these difficulties we could

anticipate and take precautions during upcoming experiments


Ho

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25

Recommendations for future research In this study we found that hypothermic oxygenated machine perfusion is superior to cold

storage when evaluating renal function during 4 hour normothermic perfusion in a pig

slaughterhouse model However we do suspect that adding mannitol dexamethason insulin

and more nutrients during reperfusion could improve kidney function More experiments

regarding the last experimental group should be performed to prove our suspicions

Conducting more experiment should make us more familiar with the perfusion techniques

allowing us to better anticipate on technical difficulties Defects in equipment could be solved

more quickly or can be prevented

Conclusions In this study we created a low cost normothermic machine perfusion DCD model with porcine

slaughterhouse kidneys A stable 4 hour pressure controlled perfusion is established with

mean pulsatile arterial pressure of 75 mmHg We were able to keep the perfusate temperature

37 degC and the partial oxygen pressure above 60 kPa After performing several experiments

regarding preservation and perfusion techniques 30 minutes of warm ischemia combined

with hypothermic oxygenated machine perfusion and additives during reperfusion seemed

superior to all other experimental groups However kidney function still remains suboptimal

compared to other studies Due to technical difficulties while performing experiments with

additives during normothermic machine perfusion in the 30WI+HMPO2+NMP+ group the

number of kidneys included in this experimental group is small Further research needs to be

conducted to determine the optimal way of delivering normothermic machine perfusion in the

reperfusion period


Ho

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26

Bibliography 1 Davis AE Mehrotra S McElroy LM et al The extent and predictors of waiting

time geographic disparity in kidney transplantation in the United States Transplantation 2014 May 2797(10)1049-57

2 Gerber LM Chiu Y Carney N et al Marked reduction in mortality in patients

with severe traumatic brain injury J Neurosurg 20131191583ndash1590

3 Maggiore U Cravedi P The marginal kidney donor Current opinion in organ

transplantation 19 (4) 372-380 (2014)

4 Paul KT Avezaat CJ Ijzermans JN et al Organ donation as transition work

Policy discourse and clinical practice in The Netherlands Health (London) 2014

Jul18(4)369-87

5 Perico N Cattaneo D Sayegh MH Remuzzi G Delayed graft function in kidney

transplantation Lancet 364 (9447) 1814-1827 (2004)

6 Kootstra G Daemen JH Oomen AP Categories of nonheartbeating donors

Transplant PRoc 1995272893-4

7 Tennankore KK Kim SJ Alwayn IP Kiberd BA Prolonged warm ischemia time

is associated with graft failure and mortality after kidney transplantation Kidney

Int 2016 89 3 648-658

8 Harper SJF Hosgood SA Nicholson ML et al The Effect of Warm Ischemic

Time on Renal Function and Injury in the Isolated Hemoperfused Kidney

Transplantation 200886 445ndash451

9 Ausania F White SA Pocock P Manas DM Kidney damage during organ

recovery in donation after circulatory death donors Data from UK National

Transplant Database Am J Transplant 2012 12932ndash936

10 Net M Valero R Almenara R et al Hepatic xanthine levels as viability predictor

of livers procured from non-heart-beating donor pigs Transplantation 2001 71

1232

11 Hosgood SA Nicholson ML Normothermic kidney preservation Curr Opin

Organ Transplant 2011 16 169

12 Johnson LB Plotkin JS Howell CD et al Successful emergency transplantation

of a liver allograft from a donor maintained on extracorporal membrane

oxygenation Transplantation 199763910-911

13 Magliocca JG Magee JC Rowe SA et al Extracorporeal support for organ

donation after cardiac death effectively expands the donor pool J Trauma 2005

58 1095-1102

14 Famey AC Singh RP Hines MH et al Experience in renal and extrarenal

transplantation with donation after cardiac death donors with selective use of

extracorporeal support J Am Coll Surg 20082061028-1037

15 Oniscu GC Randle LV Watson CJ et al In situ normothermic regional perfusion

for controlled donation after circulatory death--the United Kingdom experience

Am J Transplant 2014 Dec14(12)2846-54

16 Hessheimer AJ Billault C Barrou B Fondevila C Hypothermic or normothermic

abdominal regional perfusion in high-risk donors with extended warm ischemia

times impact on outcomes Transpl Int 2015 Jun28(6)700-7

17 Valero R Cabrer C Oppenheimer F et al Normothermic recirculation reduces

primary graft dysfunction of kidneys obtained from nonheart-beating donors

Transpl Int 2000 13303-310

18 Port FK Bragg-Gresham JL Metzger RA et al Donor characteristics associated

with reduced graft survival an approach to expanding the pool of kidney donors

Transplantation 2000232263-71


Ho

ofd

stu

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iblio

grap

hy

27

19 Demiselle J Augusto JF Videcoq M et al Transplantation of kidneys from

uncontrolled donation after circulatory determination of death comparison with

brain death donors with or without extended criteria and impact of normothermic

regional perfusionTranspl Int 2016 29 4 432-442

20 St Peter SD Imber CJ Friend PJ Liver and kidney preservation by perfusion

Lancet 2002 359604-613

21 Hosgood SA van Heurn E Nicholson ML Normothermic machine perfusion of

the kidney better conditioning and repair Transpl Int 2015 Jun28657-64

22 Yang B Hosgood SA Harper SJ Nicholson ML leucocyte depletion improves

renal function in porcine kidney hemoreperfusion through reduction of

myeloperoxidase+ cells caspase-3 IL-1 beta and tubular apoptosis J Surg Res

2010164e351

23 Gawaz M Role of platelets in coronary thrombosis and reperfusion of ischemic

myocardium Cardiovasc Res 200461498

24 Vermeulen Windsant IC Snoeijs MG Hanssen SJ et al Hemolysis is associated

with acute kidney injury during major aortic surgery Kidney Int 2010

May77(10)913-20

25 Unger V Grosse-Siestrup C Fehrenberg C et al Reference values and

physiological characterization of a specific isolated pig kidney perfusion model J

Occup Med Toxicol 2007 2 1

26 Moers C Smits JM Maathuis MH Treckmann J van Gelder F Napieralski BP et

al Machine perfusion or cold storage in deceased-donor kidney transplantation N

Engl J Med 2009360(1)7ndash19

27 Bon D Chatauret N Giraud S Thuillier R Favreau F Hauet T New strategies to

optimize kidney recovery and preservation in transplantation Nat Rev Nephrol

20128(6)339ndash47

28 Munshi R Hsu C Himmelfarb J Advances in understanding ischemic acute

kidney injury BMC Med 2011911

29 Eltzschig HK Eckle T Ischemia and reperfusion--from mechanism to

translation Nat Med 2011171391ndash1401

30 Salvadori M Rosso G Bertoni E Update on ischemia-reperfusion injury in

kidney transplantation Pathogenesis and treatment World J Transplant 2015 Jun

245(2)52-67



20128(6)339ndash47

32 Hosgood SA Barlow AD Yates PJ Snoeijs MGJ van Heurn ELW Nicholson

ML A pilot study assessing the feasibility of a short period of normothermic

preservation in an experimental model of non heart beating donor kidneys J Surg

Res 2011171(1)283ndash90

33 Nicholson ML Hosgood SA Renal transplantation after ex vivo normothermic

perfusion the first clinical study Am J Transpl 201313(5)1246ndash52

34 Moers C Pirenne J Paul A Ploeg RJ Machine Perfusion or Cold Storage in

Deceased-Donor Kidney Transplantation N Engl J Med 2012366(8)770ndash1

35 Brat A Pol RA Leuvenink HGD Novel preservation methods to increase the

quality of older kidneys Curr Opin Organ Transpl 201520(4)438ndash43

36 Nederlandse Transplanatiestichting 2015 URL

httpwwwtransplantatiestichtingnlnieuwsalle-donornieren-op-de-machine

geraadpleegd (6th July 2016)


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37 Hosgood SA Nicholson HF Nicholson ML Oxygenated kidney preservation

techniques Tranplantation 201293455

38 Thuillier R Allain G Celhay O Hebrard W Barrou B Badet L Leuvenink H

Hauet T Benefits of active oxygenation during hypothermic machine perfusion of

kidneys in a preclinical model of deceased after cardiac death donors J Surg Res

2013 Oct184(2)1174-81

39 Hosgood S Harper S Kay M Bagul A Waller H Nicholson ML Effects of

arterial pressure in an experimental isolated haemoperfused porcine kidney

preservation system Br J Surg 200693(7)879ndash84

40 Mancina E Kalenski J Paschenda P Beckers C Bleilevens C Boor P et al

Determination of the Preferred Conditions for the Isolated Perfusion of Porcine

Kidneys Eur Surg Res 201554(1-2)44ndash54

41 Bagul A Hosgood S Kaushik M Kay MD Waller HL Nicholson ML

Experimental renal preservation by normothermic resuscitation perfusion with

autologous blood Br J Surg 200895(1)111ndash8


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29

Acknowledgements This study would not have been possible without the help of all people mentioned below

First of all many thanks to my faculty supervisor prof dr Henri Leuvenink for his

inspirational support and valuable feedback on the design and process of this study

Special thanks to my daily supervisor Leonie Venema who was always available to listen to

my problems and give advice Also for always accompanying me during all our experiments

most of all during our trips to the slaughterhouse at the crack of down I am especially

grateful for all her efforts pushing me to achieve more intellectual milestones then I imagined

reaching at the start of this project

Many thanks to dr Niels lsquot Hart for all his help regarding histological evaluation and most of

all making beautiful pictures of our stainings

Also special thanks to Aukje Brat and Merel Pool for all their help with the experiments from

preparing the kidney to cleaning everything up It would not have been possible without their

support

Last but not least many thanks to Jacco Zwaagstra Wendy Oost employees of the UMCG

surgical research department employees of slaughterhouse Hilbrants and Kroon and to all the

others who were otherwise involved


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30

Appendix 1 Protocol for organ and blood retrieval

Slaughterhouse kidneys and blood

Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel

o 5ml25000 IE Heparine

o 5ml syringe with needle

- Kidneys (depending on the manner of transportation)

o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage

Transport box with crushed ice

o Hypothermic machine perfusion

Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable

Canularsquos and patch

holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice

o Subnormothermic machine perfusion

Kidney assist + sensors+ batteries

Oxygen bottle if needed

KA Disposable adapted to fit the canula used for NMP

Oxygen bottle

Canula for artery

Cannula for urether

Sutures to secure cannula

Sutures to repair leakage if necessary

20 ml syringe

Blunt needle

Heat packs (place them in a 37degC incubator the night before)

500 ml Ringerslactate 37degC

Nacl 37degC

500 ml beaker

Scale

Protocol

Blood

- Put the Heparine in the 5L beaker with the syringe

- Catch about 3 liter blood with the beaker

- Poor the blood in a jerry can use a funnel if needed

Kidneys

- Fill the styrofoam box with ice and place the gauze on top Use a syringe and some

NaCL to wet the gauze Place the kidney pair on top with the aorta facing upwards

When the preferred ischemic time is passed Important in case of sNMP donrsquot use ice

or cold fluids

- Cut the aorta lengthwise in half and search the renal arteries Make sure you donrsquot

damage the renal arteries

- Fill the 60 ml syringe with cold NaCl and attach the catheter

- Place the catheter carefully in the renal artery and flush slowly Be careful not to apply

excessive pressure with the syringe Keep flushing until the kidneyrsquos aspect had

become uniformly pale and clear fluid runs from the vena

- Remove the catheter

- Remove the contra lateral kidney

- Store the kidney for transport

o Cold storage

Place the kidney in a organ bag with cold NaCl

Place this bag in a larger bag containing ice

Place the bag in a large transport box filled with ice


While one person is taking care of the kidney flush another person has

to assemble and prime the Kidney Assist using UW-machine perfusion

Fill the Kidney Assist transport box with ice Donrsquot forget to open the

oxygen bottle if needed


Ch

apte

r S

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ghte

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ki

dn

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and

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od

32


After flushing the kidney remove excessive fat from the kidney except

near the urether and hilum Connect the aorta patch to the patch holder

Use an artificial cannula if needed Place the patch holder in the kidney

holder check for leakage with a 20ml syringe



Ch

apte

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rho

use

ki

dn

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and

blo

od

33


Place the kidney holder inside the kidney assist reservoir and start

perfusion

Take a sample off the perfusate after 15 ml of perfusion and write

perfusion parameters down on the CRF



to assemble and prime the Kidney Assist using 500ml warm ringers

lacate and 500ml whole blood Fill the Kidney Assist transport box

with the heatpacks Donrsquot forget to turn the oxygen bottle open

Once the kidney is flushed weigh the kidney and write it down

Remove all excessive fat from the kidney except near the urether and

hilum

Place the cannula in the renal artery and secure it with a suture Check

for leakage with a syringe

Place a cannula in the urether and secure it with a suture check for

leakage and correct placement with a bolus of warm NaCl by using

syringe and blunt needle

Place the kidney in the reservoir and start perfusion

Take a sample off the perfusate after 15 min of perfusion and write

perfusion parameters down

During the whole procedure note the following time points

- Time of death of the pig start warm ischemia

- Moment of starting flush end warm ischemia

- Moment were transportation starts start cold ischemia

- In case of HMP or sNMP take a sample at 15 minutes of perfusion and the end of

perfusion Also note the hemodynamics


Ch

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34

Preparations at the lab

Leukocyte depleted blood

Materials

- Catheter bag

- Funnel with silicone tubing to connect to catheter bag

- Clamps

- Jerrycan filled with blood at the slaughterhouse

- Leukocyte filter (BioR O2 plus BS PF Fresenius Kabi Bad Homburg Germany)

- 2L beaker

Protocol

- Fill the catheter bag with blood using the funnel

- Close the inlet with a clamp

- Attach the leukocyte filter to the outlet off the catheter bag

- Hang the system to a hook an place the beaker underneath

- Open the outlet of the catheter bag and de-air the leukocyte filter place the beaker

underneath NB Make sure you keep an eye on the beaker there is always a risk of

overflow

A blood sample is analysed for Hematocrit and white blood cell count before blood enters the

NMP system


Ch

apte

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orm

oth

erm

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nal

per

fusi

on

cir

cuit

35

Appendix 2 Protocol NMP

Normothermic regional perfusion circuit

Materials Cabinet with heater and thermostat

Kidney Assist (Organ Assist Groningen Netherlands) with heart assist software

Centrifugal pumphead (Deltastream DP3 MEDOS Medizintechnik AG Stolberg Germany)

Pressure sensor (Truewave disposable pressure transducer Edwards lifesciences Irvine

California USA)

Temperature sensor

Clamp-on flow sensor (ME7PXL clamp Transonic Systems Inc Ithaca NY)

Oxygenator with integrated heat exchanger (HILITE 1000 MEDOS Medizintechnik AG

Stolberg Germany)

Orgaan chamber

Arterial canula without patch (cannula for organ perfusion ndash 12F INFUSION Warszawa)

Waterbath

Luer Lock T- connector 14-14

Luer Lock T-connector 316-316

Connector 14-38

14 silicone tubing ndash 40 cm (2x)

14 silicone tubing ndash 15cm

14 PVC tubing ndash 35 cm


14 PVC tubing - 60 cm

38 PVC tubing ndash30 cm

ndash 30 cm

Luer Lock three-way valves (2x)

Infusion tubing (2x)

Tie wraps


Ch

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36

Assembling the system Connect all the components above

- Connect the outflow of the pump head to the blood inlet of the oxygenator with 30 of

38 frac14 PVC tubing

- The blood outlet of the oxygenator is connected to a luer lock connector ( frac14 - frac14 ) with

10 cm silicon tubing The pressure sensor is connected to the luer lock connector with

the infusion tubing

- The other outlet on the oxygenator is connected to the infusion tube with at the and a

luer lock valve

- Connect 10 cm frac14 PVC tubing from the luer lock connecter with the pressure sensor to

the inlet of the organ chamber

- The outlet of the organ chamber is connected with a frac14 -38 connector A 30 cm 38

PVC tube is then attached and connected to the inlet of the pumphead

- The water bath is connected to the in- and outlet of the water compartment in the

oxygenator with 40 cm frac14 silicone tubing NB pay attention to the water flow the

outflow of the water bath should be connected to the inlet of the oxygenator

- Connect 60 cm frac14 PVC tubing to the oxygen inlet on the oxygenator and attach the

other end to the carbogen supply

- The temperature sensor floats in the organ chamber

- The clamp-on flow sensor is attached to the frac14 silicone tubing leading from the outlet

of the oxygenator to the luer lock connector with the pressure sensor It is optional to

use Vaseline to improve signal transduction

- Make sure every connection is tie wrapped to avoid leakage under pressure

Figure 2 Perfusion circuit


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37

Normothermic machine perfusion

Materials

- Assembled perfusion ciruit (See lsquonormothermic machine perfusion circuitrsquo)

- 300ml Ringerslactate

- 10ml Voluven

- 8ml 84 Natrium bicarbonate

- 90 mg Creatinine

- 100mg200mg Augmentin

- 20mgml Sodium nitroprusside (100microl of the dilution)500 ml leukocyte depleted

blood (See lsquoleukocyte depleted bloodrsquo)

Protocol

- Fill the water bath with purified water and set the temperature at 38degC

- Set the temperature off the external thermostat at 37degC this will regulate the

temperature inside the cabinet Place the temperature probe in the cabinet and turn the

heater in the cabinet on

- Prime the system with the priming fluid

o 300ml Ringerslactate

o 10ml Voluven

o 8ml 84 Natrium bicarbonate

o 90 mg Creatinine

o 100mg200mg Augmentin

o 100microl Sodium nitroprusside

- De-air the tubing leading from the organ chamber to the pump head passively Then

attach the pump head to the Kidney Assist pump unit

- Turn on the external flow unit

- Attach the pressure sensor temperature sensor and flow sensor to the pump unit

- Power on the kidney assist and follow the priming menu

o Press lsquopowerrsquo button

o lsquoSelftest OKrsquo press push-dial button

o ldquodisposable connectedrdquo press push-dial button

o ldquoPerfusate level OKrdquo press push-dial button

o In priming mode remove air from oxygenator bubble trap by opening the

valve on top of oxygenator Close valve once air is removed

o Remove air from infusion lines

o Turn valve on pressure sensor in direction of the perfusion circuit remove caps

on the pressure sensor Pull bleu snap tap and use a syringe to aspirate the

perfusate until a few drops drip out

o Press push-dial button to calibrate the pressure sensor

o Replace the caps on the pressure sensor and turn the valve in direction of the

side port

o press push-dial button and set pressure on 75 mmHg

o Stop when ldquoconnect heartrdquo shows on the display

- Open the carbogen source and set the flow regulator at 05 mlmin

- Add 500 ml leukocyte depleted blood

- Wait until the priming solution reaches 37degC before connecting the kidney

- Meanwhile prepare the kidney for perfusion


Ch

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38

Preparing the kidney

Materials

- Icebox with crushed ice

- Large gauze (40x40)

- Syringe 60 ml and 20 ml

- Blunt needle

- Artery cannula

- Urether cannula

- Surgical instruments

- Biopsy gun

- 4 Formalin + biopsy holder and gauze

- SONOP

- Liquid nitrogen

- scale

Protocol

- Place kidney on wet gauze with crushed ice

underneath

- Remove all excessive fat from the kidney except near

the urether and hilum

- Place a cannula inside the urether and tie 2-0 braided

suture around distal end of urether to make sure it

remains in the same place Check for leakage and

correct placement with a bolus of NaCl by using a


- Place a cannula inside the renal artery secure it with a

suture and check for leakage using a syringe

- Weigh the kidney and write it down

- Take a biopsy using the biopsy gun Store one half in

formalin store the other half in SONOP in liquid

nitrogen

Perfusion

To start perfusion

- Place the prepared kidney in the organ chamber

- Check if the system is still free of air bubbles If not remove them

- Connect the artery cannula to the perfusion circuit make sure to keep the system air

free

- Press push-dial button to start perfusion

- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes

Figure 7 Cannulated kidney


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39

- Infusion caps

- Beaker

- Crushed ice in a styrofoam box

- CRF

- Warm ringers lactate (place within the heat cabinet)

- 5 Glucose

- 5ml tubes

Protocol

- Place a beaker under the cannula of the ureter Make sure that the distal tip of this

cannula is below the level of the renal pyelum

- Write down the start time and hemodynamics on the CRF

- Take samples on given time points 05 ml from sample line and 05 ml from the vena

and analyse immediately using the ABL800 bloodgas analyser Store 5ml perfusate

drawn from the sample line on ice Before taking the sample draw some perfusate

from the sample line to remove death volume

- Replace the beaker underneath the cannula of the ureter at the correct time points

Store urine on ice

- Replace the sample and urine volume using the sample line 6ml for the samples + the

amount of urine collected

- Check the glucose concentration on the bloodgas results If the number drops below 8

mmolL add glucose according to the scheme

Figure 8 Kidney connected to NMP circuit


Ch

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40

Cleaning the system

Materials

- 4 formalin + biopsy holder

- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol

- Shut down the Kidney Assist using the power button

- Disconnect the kidney and weigh it

- Take 2 large biopsies store 1 in formalin Place the other on a filtration paper and

place it on a petridish Then drop de petridish on the liquid nitrogen with the biopsy on

the upper side

- Discard the kidney following regulations

- Close the carbogen and disconnect tubing leading to the oxygenator

- Disconnect al sensors carefully

- Turn off heater inside cabinet (and external thermostat)

- Turn off the water bath and disconnect tubing leading to the oxygenator

- Remove the pump from the Kidney Assist

- Remove the perfusion circuit from the cabinet and disconnect Rinse tubing with

plenty of purified water until the tubing appears clean Then rinse it with more purified

water Rinse the oxygenator with plenty of purified water

- Dry the system and oxygenator using carbogen

- Clean cabinet with ethanol and close it

- Check if the area surrounding the experimental set up is clean

- Store all samples in -80 degC after 10 minutes centrifugation at 4degC 1000g Except for

the formalin biopsies they must be embedded in paraffin wax immediately


Ch

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41

Appendix 3 Results overview




2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

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42



Graph 6amp4 Mean Renal blood flow in mlmin100 gram per experimental group

40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

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the

syst

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43



Graph 9 Mean Renal blood flow in mlmin100 gram per experimental group

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


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44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930

` Graph 10 Mean urine production in mlmin per experimental group



15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


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45

Table 6 Post Hoc analysis of urine production per time point




120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

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46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

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47

Table 11 Mean values and standard deviation of kidney function and tissue injury parameters

Group Creatinine clearance (mlmin100gr) AUC

Edema FeNa+ () AUC

30WI+CS 24 (plusmn2794) 12 (plusmn 14) 16317 (plusmn186)

7WI+CS 578 (plusmn21953) 30 (plusmn01) 7309 (plusmn889)


40W+sNMP 160 (plusmn19814) 27 (plusmn14) 3896 (plusmn54)

30WI+HMP+O2 383 (plusmn25962) 1367 (plusmn 305) 7346 (plusmn6240)

30WI+HMP-O2 354 (plusmn285) 13 (plusmn848) 8685 (plusmn2356

30WI+HMP+O2+NMP+ 1250 15 3134


Group LDH concentration (UL) AUC

Serum Creatinine drop ()

Urine production accumulated (ml)

30WI+CS 172250 (plusmn 132080) 15 (plusmn 54) 61 (plusmn39)



40W+sNMP 111120 (plusmn 5441) 12 (plusmn 14) 33 (plusmn6)

30WI+HMP+O2 261603 (plusmn 203398) 67 (plusmn 249) 211 (plusmn112)

30WI+HMP-O2 143748 (plusmn 1118) 645 (plusmn 2333) 493 (plusmn447)

30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027




Table 14 Post Hoc analysis of kidney function parameters

Parameter 30WI+CS vs 30WI+HMP+O2



FeNa+ () AUC 0032 0006 0589

Serum creatinine drop ()

0007 0001 0436


Ch

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48

Table 15 statistical analysis of pH per time point


-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)

Lactate dehydrogenase

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2 30WI+HMP-O2


Ch

apte

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49

Table 16 statistical analysis of LDH per time point


15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075

Table 17 statistical analysis of lactate per time point


15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

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ract

sam

enva

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2

Abstractsamenvatting Objectives To decrease waiting time for a kidney transplant donor kidneys of inferior quality

are increasingly being accepted Normothermic regional perfusion (NRP) in donation after

circulatory death restores abdominal circulation with autologous blood and improves organ

quality The use of blood in machine perfusion can cause inflammation injury tissues damage

and cell death Probably it is better to replace the blood in NRP with an artificial preservation

solution To test the different solutions a normothermic machine perfusion model for an

isolated kidney has to be designed The aim of this study is to design a NMP model with

porcine slaughterhouse kidneys to test kidney viability

Methods Porcine kidneys and autologous whole blood were obtained from the

slaughterhouse Kidneys were transported to our lab and reperfused for 4 hours using the

NMP set-up Seven groups were created (n=1-4) Kidneys were transported differently using

cold storage subnormothermic machineperfusion oxygenated- or non-oxygenated

hypothermic machine perfusion Warm and cold ischeamic times differ between groups In

one group mannitol insulin nutrients and dexamethson was added during NMP (NMP+)

Hemodynamics were monitored and perfusate and urine samples were taken regularly

Biopsies were taken to asses renal histology

Results A stable NMP was established There was a significant difference between groups

Kidneys preserved using oxygenated hypothermic machine perfusion and reperfused with

NMP+ performed significantly better These kidneys showed better creatinine clearance

sodium reabsorption and urine production than the control group

Conclusion The model created using oxygenated hypothermic machine perfusion and NMP+

likely is useful for testing different perfusion solutions However more research is required to

optimize this model

Doelstellingen Om wachttijd voor een niertransplantatie te verkorten worden donornieren

van mindere kwaliteit in toenemende mate geaccepteerd Normotherme regionale perfusie

(NRP) na donatie na hartdood hersteld de abdominale circulatie met autoloog bloed en

verbeterd de orgaan kwaliteit Het gebruik van bloed in machine perfusie kan inflammatoire

schade weefselschade en celdood veroorzaken Waarschijnlijk is het beter om bloed

tevervangen door een artificieumlle preservatie vloeistof Om verschillende vloeistoffen te

kunnen testen moet er een normotherm machineperfusie (NMP) model ontworpen worden

Het doel van deze studie is het ontwerpen van een NMP model met varkens nieren uit het

slachthuis om de kwaliteit van de nieren te testen

Methode Varkens nieren en autoloog bloed werden verzameld in het slachthuis De nieren

werden vervoerd naar ons lab en 4 uur op de NMP opstelling geperfundeerd 7 groepen zijn

getest met 1-4 nieren per groep Er zijn verschillende manieren van transport gebruikt cold

storage subnormotherme machineperfusie geoxygeneerde of niet ge-oxygeneerde

hypotherme machineperfusie Warme en koude ischemie tijden verschillen per groep In een

groep is tijdens NMP mannitol insuline voedingsstoffen en dexamethason toegevoegd

(NMP+) De hemodynamica werd gemonitord en perfusaat en urine monsters werden

regelmatig genomen Biopsieeumln werden genomen om renale histologie te beoordelen

Resultaten Een stabiel NMP systeem werd verkregen De groep waarbij geoxygeneerde

hypotherme machineperfusie en NMP+ gebruikt werd presteerde significant beter dan de

controle groep Er was betere creatinine klaring natrium reabsorptie en urine productie

Conclusie Het gecreeumlerde model waarbij gebruik gemaakt wordt van geoxygeneerde

hypotherme machineperfusie en NMP+ is bruibaar om verschillende vloeistoffen voor

perfusiedoeleinden te testen Verder onderzoek is nodig om de nierfunctie tijden de NMP

periode te verbeteren


Ho

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sam

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3


Introduction 4

Organ shortage 4



Machine perfusion 7

Study objectives 8




Kidney transport 10

Perfusion 11



Results 14



Renal function 18

Renal Histology 22

Discussion 23

Considerations 23



Bibliography 26

Acknowledgements 29

Appendices 30


Ho

ofd

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tro

du

ctio

n

4

Introduction






























United states

(National Kidney

Foundation)


(Eurotransplant


statistics report)

Patients on kidney


100791 (January 2016) 10282 (March 2016)


transplanted

11570 (2014) 1827 (2015)



transplant



Ho

ofd

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k In

tro

du

ctio

n

5






















NRP has been













Maintaining







or hypertension18






Furthermore




Ho

ofd

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tro

du

ctio

n

6


























injury22






















Ho

ofd

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tro

du

ctio

n

7



ischemia






assessed25











transplantation26









IRI is principally






in the recipient29










preservation21



NMP may be able to



Kidneys that


Ho

ofd

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tro

du

ctio

n

8


cold storage33

























animal model













Ho

ofd

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ater

ial a

nd

met

ho

ds

9


Experimental design














perfusion












experimental groups




2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

10














Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

11


























Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

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k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k A

bst

ract

sam

enva

ttin

g

3


Introduction 4

Organ shortage 4



Machine perfusion 7

Study objectives 8




Kidney transport 10

Perfusion 11



Results 14



Renal function 18

Renal Histology 22

Discussion 23

Considerations 23



Bibliography 26

Acknowledgements 29

Appendices 30


Ho

ofd

stu

k In

tro

du

ctio

n

4

Introduction






























United states

(National Kidney

Foundation)


(Eurotransplant


statistics report)

Patients on kidney


100791 (January 2016) 10282 (March 2016)


transplanted

11570 (2014) 1827 (2015)



transplant



Ho

ofd

stu

k In

tro

du

ctio

n

5






















NRP has been













Maintaining







or hypertension18






Furthermore




Ho

ofd

stu

k In

tro

du

ctio

n

6


























injury22






















Ho

ofd

stu

k In

tro

du

ctio

n

7



ischemia






assessed25











transplantation26









IRI is principally






in the recipient29










preservation21



NMP may be able to



Kidneys that


Ho

ofd

stu

k In

tro

du

ctio

n

8


cold storage33

























animal model













Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

9


Experimental design














perfusion












experimental groups




2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

10














Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

11


























Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k In

tro

du

ctio

n

4

Introduction






























United states

(National Kidney

Foundation)


(Eurotransplant


statistics report)

Patients on kidney


100791 (January 2016) 10282 (March 2016)


transplanted

11570 (2014) 1827 (2015)



transplant



Ho

ofd

stu

k In

tro

du

ctio

n

5






















NRP has been













Maintaining







or hypertension18






Furthermore




Ho

ofd

stu

k In

tro

du

ctio

n

6


























injury22






















Ho

ofd

stu

k In

tro

du

ctio

n

7



ischemia






assessed25











transplantation26









IRI is principally






in the recipient29










preservation21



NMP may be able to



Kidneys that


Ho

ofd

stu

k In

tro

du

ctio

n

8


cold storage33

























animal model













Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

9


Experimental design














perfusion












experimental groups




2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

10














Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

11


























Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k In

tro

du

ctio

n

5






















NRP has been













Maintaining







or hypertension18






Furthermore




Ho

ofd

stu

k In

tro

du

ctio

n

6


























injury22






















Ho

ofd

stu

k In

tro

du

ctio

n

7



ischemia






assessed25











transplantation26









IRI is principally






in the recipient29










preservation21



NMP may be able to



Kidneys that


Ho

ofd

stu

k In

tro

du

ctio

n

8


cold storage33

























animal model













Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

9


Experimental design














perfusion












experimental groups




2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

10














Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

11


























Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k In

tro

du

ctio

n

6


























injury22






















Ho

ofd

stu

k In

tro

du

ctio

n

7



ischemia






assessed25











transplantation26









IRI is principally






in the recipient29










preservation21



NMP may be able to



Kidneys that


Ho

ofd

stu

k In

tro

du

ctio

n

8


cold storage33

























animal model













Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

9


Experimental design














perfusion












experimental groups




2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

10














Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

11


























Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k In

tro

du

ctio

n

7



ischemia






assessed25











transplantation26









IRI is principally






in the recipient29










preservation21



NMP may be able to



Kidneys that


Ho

ofd

stu

k In

tro

du

ctio

n

8


cold storage33

























animal model













Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

9


Experimental design














perfusion












experimental groups




2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

10














Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

11


























Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k In

tro

du

ctio

n

8


cold storage33

























animal model













Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

9


Experimental design














perfusion












experimental groups




2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

10














Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

11


























Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

9


Experimental design














perfusion












experimental groups




2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

10














Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

11


























Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

10














Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

11


























Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

11


























Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

12
















Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k M

ater

ial a

nd

met

ho

ds

13
























necessary


Perfusion

parameters



levels

GFR LDH


excretion

Lactate


Kidney weight ATP


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

14

Results















Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

15




above 60 kPa










at t=0


40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

16









kidney








Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

17





0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

18











15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

19





table 9








0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+













Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

20





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165



















0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k R

esu

lts

21


two groups























90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548




Edema 0279

FeNa+ () AUC 0027





Ho

ofd

stu

k R

esu

lts

22







in these groups













groups



Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

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esu

lts

22







in these groups













groups



Ho

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n

23











As we

























group












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24















































Ho

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25




















reperfusion period


Ho

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hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

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2013 Oct184(2)1174-81











Ho

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stu

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ow

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gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k D

iscu

ssio

n

23











As we

























group












Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k D

iscu

ssio

n

24















































Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k D

iscu

ssio

n

25




















reperfusion period


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

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stu

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gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k B

iblio

grap

hy

26









Jul18(4)369-87







Int 2016 89 3 648-658









1232








58 1095-1102












Transpl Int 2000 13303-310





Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k B

iblio

grap

hy

27






Lancet 2002 359604-613






2010164e351





May77(10)913-20









20128(6)339ndash47







245(2)52-67



20128(6)339ndash47




Res 2011171(1)283ndash90











Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k B

iblio

grap

hy

28






2013 Oct184(2)1174-81











Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k A

ckn

ow

led

gem

ents

29













support





Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ho

ofd

stu

k S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

30



Materials

- Blood collection

o 5L beaker

o Jerrycan

o Funnel




o General supplies

1L NaCl for flush

Surgical scissors

(sharp) 2x

Surgical forceps 3x

Clamps

Syringe 60 ML with

tip

Catheter (5cm) for

flush

Large gauze

(40x40cm)

Styrofoam box for

inspecting the

kidneys

Gloves

Trash bags

Pen + paper

o Cold storage

Organ bags

NaCL for storage



Kidney assist +

sensors+ batteries

Oxygen bottle if

needed

KA Disposable


holder

UW- machine

perfusion solution

Sutures

20 ml syringe

Crushed ice





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





Oxygen bottle

Canula for artery

Cannula for urether



20 ml syringe

Blunt needle



Nacl 37degC

500 ml beaker

Scale

Protocol

Blood




Kidneys




or cold fluids










o Cold storage










Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

32








Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r S

lau

ghte

rho

use

ki

dn

eys

and

blo

od

33



perfusion










hilum
















Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r P

rep

arat

ion

s at

th

e la

b

34



Materials

- Catheter bag


- Clamps



- 2L beaker

Protocol







overflow


NMP system


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

35







California USA)

Temperature sensor



Stolberg Germany)

Orgaan chamber


Waterbath



Connector 14-38







ndash 30 cm



Tie wraps


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

36






the infusion tubing


luer lock valve

















Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r N

orm

oth

erm

ic r

egio

nal

per

fusi

on

cir

cuit

37


Materials



- 10ml Voluven


- 90 mg Creatinine




Protocol







o 10ml Voluven


o 90 mg Creatinine




















side port








Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r P

erfu

sio

n

38


Materials




- Blunt needle

- Artery cannula

- Urether cannula


- Biopsy gun


- SONOP

- Liquid nitrogen

- scale

Protocol


underneath













nitrogen

Perfusion

To start perfusion




free


- Close the cabinet

During perfusion

Materials

- 1ml syringes

- 5 ml syringes

- 10 ml syringes



Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r P

erfu

sio

n

39

- Infusion caps

- Beaker


- CRF


- 5 Glucose

- 5ml tubes

Protocol









Store urine on ice







Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

sys

tem

40

Cleaning the system

Materials


- Liquid nitrogen

- Filtration paper

- petridish

- Biotex

- Purified water

- Blade (mesje)

- ethanol

Protocol





the upper side
















Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

41





2 2 78 CS 2 NMP

3 2 20 CS 3 NMP

4 2 40 sNMP 2 NMP

5 4 30 HMP + O2 2-3 NMP

6 2 30 HMP - O2 3 NMP

7 2 30 HMP + O2 3 NMP +




Kidney 1 Kidney 2


t=0 351 53 341 40

t=15 36 51 354 35 4

t=30 384 87 369 120 5

t=60 37 215 6 369 145 10

t=90 362 230 37 130 105

t=120 363 194 6 366 94 55

t=150 371 175 353 38 6

t=180 369 173 5 361 48 4

t=210 375 180 36 74 65

t=240 37 194 11 364 85 105

Total 28 Total 62



t=0 323 25

t=15 339 156 35

t=30 367 196 9

t=60 373 204 17

t=90 372 205 115

t=120 371 205 15

t=150 374 204 16

t=180 36 190 12

t=210 371 184 105

t=240 373 183 8

Total 1025


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

42




40

45

50

55

60

65

70

75

80

-10 15 30 60 90 120 150 180 210 240

kPa

Time

pO2

Kidney 1

Kidney 2

Kidney 4

0

2

4

6

8

10

12

-10 15 30 60 90 120 150 180 210 240

mm

ol

L

Time

Glucose

kidney 1

kidney 2

kidney 4

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+CS

0

50

100

150

200

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

7WI+CS


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

43




0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

20WI+CS

0

20

40

60

80

100

120

140

160

0

15

3

0

50

7

0

90

1

10

1

30

1

50

1

70

1

90

2

10

2

30

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

40WI+sNMP

0

20

40

60

80

100

120

140

160

0 20 50 80 110 140 170 200 230

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2

0

20

40

60

80

100

120

140

160 0

15

3

0

50

70

9

0

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP-O2

0

20

40

60

80

100

120

140

160

0

15

30

50

70

90

11

0

13

0

15

0

17

0

19

0

21

0

23

0

flo

w (

ml

min

10

0gr

)

Time (min)

Flow

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

44




0 0063 120 0833

10 0135 130 0954

15 0372 140 0971

20 0586 150 0986

30 0743 160 0997

40 0743 170 0998

50 0777 180 0998

60 0759 190 0998

70 0798 200 0997

80 0723 210 0991

90 0714 220 0979

100 0852 230 0968

110 0846 240 0930




15 0066

30 0163

60 0071

90 0065

120 0039

150 0050

180 0110

210 0078

240 0079

0

1

2

3

4

5

6

7

15 30 60 90 120 150 180 210 240

uri

ne

pro

du

ctio

n (

ml

min

10

0gr

)

Time point (min)

Urine production

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

45





120 0062 0001 0008

150 0065 0002 0025


0

1

2

3

4

5

6

7

8

15 30 60 90 120 150 180 210 240

cle

aran

ce r

ate

(m

lm

in1

00

gr)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+





15 0645 0004 0001

90 0028 0000 0034

120 0043 0000 0157

180 0074 0000 0137

210 0052 0000 0165













Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

46


0

20

40

60

80

100

120

140

30 60 90 120 150 180 210 240

Frac

tio

nal

exr

eti

on

of

sod

ium

(

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+















90 0001 0029 0518

120 0001 0033 0189

150 0000 0012 0653

180 0000 0015 0876

210 0001 0019 0657

240 0000 0020 0548


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

47



Edema FeNa+ () AUC







30WI+HMP+O2+NMP+ 1250 15 3134











30WI+HMP+O2+NMP+ 348465 92 812




Edema 0279

FeNa+ () AUC 0027








FeNa+ () AUC 0032 0006 0589


0007 0001 0436


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

48



-10 0836

15 0993

30 0953

60 0690

90 0349

120 0382

150 0237

180 0273

210 0168

240 0187

7

71

72

73

74

75

76

77

78

79

8

-10 15 30 60 90 120 150 180 210 240

pH

Time point (min)

pH 30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

0

500

1000

1500

2000

2500

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μL

)

Time point (min)


30WI+CS

7WI+CS

20WI+CS

40WI+sNMP



Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+


Ch

apte

r C

lean

ing

the

syst

em

49



15 0066

30 0061

60 0070

90 0098

120 0074

150 0079

180 0076

210 0070

240 0075



15 0438

30 0664

60 0647

90 0524

120 0596

150 0571

180 0501

210 0306

240 0737

10

12

14

16

18

20

22

24

26

15 30 60 90 120 150 180 210 240

con

cen

trat

ion

(μ

L)

Time point (min)

Lactate

30WI+CS

7WI+CS

20WI+CS

40WI+sNMP

30WI+HMP+O2

30WI+HMP-O2

30WI+HMP+O2+NMP+

developing a porcine slaughterhouse model for normothermic...

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