role of blood bank in stem cells therapy

8/8/2019 Role of Blood Bank in Stem Cells Therapy

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Role of blood bank in stem cells

therapy

Auda S. Aziz

Dharmais Cancer Hospital


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Topics

Blood bank activities

Collection of stem cells by apheresis Stem cells preservation

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Dharmais Blood bank activities

3 groups of activities:

Blood component services

Apheresis services

Stem cells preservation

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Blood component services

Receive stock of screened-blood componentsfrom the IRC

Store blood components Pre-transfusion testing

Patients ABO & Rhesus blood grouping Confirmation of donors ABO & Rhesus blood

group Crossmatching of donors & patients blood Issuing the compatible blood components to the

patient

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Apheresis services

Apheresis for donors blood component Screening the blood donor Performing apheresis for required blood

component, esp. Platelet Concentrate

Therapeutic Apheresis Screening the patients

Performing thrombapheresis, leukapheresis,plasmapheresis or apheresis for red cellsaccording to doctors requirement

Apheresis for stem cells (PBSCs)

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Stem cells preservation

Collection of MNCs

Cryopreservation of MNCs Thawing the MNCs before infusion

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Role of blood bank in stem cells therapy

1. Collects the hematopoietic peripheral blood

stem cells (PBSCs) by apheresis2. Preservation of hematopoietic stem cells:

Bone marrow derived hematopoietic stem cells

Peripheral blood hematopoietic stem cells

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PBSCs collection by apheresis

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Eligibility of patients / donors

Autologous

Patients eligibility for the apheresis procedureAllogeneic

Donors physical ability for the procedures

The willingness of the donors

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Patients / donors selection Selection criteria

Minimal Hb level : 8 g / dl

Platelet count > 150,000 / ul

Acceptable conditions to undergo themobilization and leukapheresis procedure

Non reactive for transfusion transmittedinfections (for allogeneic transplant )

Acceptable CD34+ pre-count

Informed consent

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Mobilizing the stem cells

Mobilizing the hematopoietic stem cells from

marrow compartment into the peripheralblood prior to collection

Using hematopoietic growth factors G-CSF5 10 ug / kg subcutaneus, for 5 days

For malignant blood diseases : G-CSF 5 10 ug / kg with chemotherapy

G-CSF alone has a poor yield in PBSCscollection

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CD34+ cells pre-count

WBC count alone does not predict the CD34+cells yield

Peripheral blood CD34+ cell count corralateswith the final cell yield

No correlation between the pre-apheresisCD34+ cell number and the percent CD34+

efficiency It is well known that the number of cycles of

chemotherapy / inversely affects the collectionefficiency and yields of CD34+ cells

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Day 5 is an appropriate time to start apheresis

If not sufficient, the second apheresis on day 6could be performed (G-CSF still given to patienton day 5 )

A minimal of pre-apheresis CD34+ cells

concentration 30 / ul can optimize yield andmaximize cost effectiveness

Collecting the stem cells

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Peripheral blood CD34+ cells concentration

exceeded 40 / ul more than 2.0 x 106

CD34+/ kgBW could be expected by a single apheresis

Absolute efficiency in allogeneic PBSCs donors:35 % - 45.5 %


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Prophylactic oral calcium given prior toapheresis procedure

Collection performed via peripheral or centralvenous

Using apheresis machine:

Continuous flow centrifugation, or

Intermittent flow centrifugation


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Continuous flow centrifugation

Cobe Spectra

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Intermittent flow centrifugation

MCS+

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For autologous transplant

Minimal threshold of CD34+ : 0.75 1 x 106 / kg

Recent data support an economic benefit associated

with greater CD34+ cell collections : 5.0 x 106 / kg (accelerated platelet engraftment, reduced febrilecomplications and use of antibiotics after transplant )

For allogeneic transplant

Minimum dose of 2.0 x 106 / kg of CD34+ cells For stem cells therapy

Minimal dose of 25 x 106 of CD34+ cells (AMI)

Targets for stem cells collection

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Preservation of hematopoietic stemcells

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Cells selection

Collection of MNCs by adding the ficol solution

Using cobe 2991 ( for stem cells transplant ) Tube method ( for stem cells therapy )

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Cobe 2991 Is the automated cell washing machine

Used manually to isolate the BM-MNCs by ficolldensity separation and washing procedure

Needs peristaltic pump (40 ml / minute) to addthe BM into the spinning ficoll solution

Input 550 ml, output 100 ml

Ficol + MNCs

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Tube method The manually isolation of BM-MNCs by ficoll

density separation and washing procedure

Using sterile consummables, performed inbiological safety cabinet

The volume of BM processed 50 ml

The final product volume 4 10 ml

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Cryopreservation Cryoprotectant preparation :

20 % DMSO

20 % autologus plasma In normal saline

Incubate stem cells and cryoprotectant at 4 Cfor at least 15 min

Slowly mixing the same volume of stem cells andcryoprotectant

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Controlled rate freezing

Minus 1 C / min from 4 C to minus 40 C

Minus 5 C / min to minus 160 C Store in liquid nitrogen ( minus 196 C )

Cryopreservation

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Evaluation

Cell count

Total nucleated cell concentration

Mononuclear cell concentration CD34+ cells , etc

Sterility test (aerobic, anaerobic)

Cells culture

Samples : Starting materials

Final product

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Thawing

Perform near patient ward

Immediately infused to the patient

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Thank you

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role of blood bank in stem cells therapy

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