1

Autologous Liver Cell Transplantation

A new approach to liver disease therapy

Presentation at ESAO Conference, September 6th 2007, KremsProf. Toni Lindl,

(I.A.Z. München as a Partner of HumanAutoCell GmbH)

2

Market and Application Area

Preclinical Development

Early Clinical Results

Content

3

Liver Disease Problem

There are currently very limited options to treat the continuous degradation of liver function in cirrhotic patients.

While liver transplantation today is widely practiced, there is a shortage of organ supply so that many patients do not survive the waiting period for an organ.

Furthermore liver transplantation is highly expensive and requires continuous follow-on treatment.

There is a strong need for a complementary treatment for liver disease that can be applied widely

4

Content

Market and Application Area

Preclinical Development

Early Clinical Results

5

Dr. Matthias Kaufmann a was member of the Harvard laboratory of Prof Vacanti focusing on scaffold-assisted tissue engineering

First trials with liver cells took take place in Harvard in the late 80ies/ early 90ies

Dr. Kaufmann succeeds in achieving proliferation of normal (non-cancerous) rat hepatocytes in vivo

Discovered optimal proliferation by co-transplantation with of langerhans islets and hepatocytes

Development of liver assist device based upon a tissue engineering technology

6

Preclinical Studies

Lewis and Gunn rats provide a suitable in vivo model to show rescue of liver function

Hepatocyes from Lewis rats are able to conjugate bilirubin via glycosyltransferase reactions in contrary to Gunn rats.

Hepatocytes from Lewis rats were isolated and cultured on a polymer scaffold and subsequently implanted into the mesenterial region of the Gunn rats.

The implanted Gunn rats showed restoration of bilirubin conjugation typically after three to four weeks.

It was shown that the implants were accepted and vascularized by the host.

Kaufmann et al. (1994) Transplant Proc 26: 2240

Kaufmann et al. (1999) Pediatr. Surg Int 15:168

7

HAC‘s scaffold design and function

The scaffold‘s material and its form is of a specific 3-dimensional structure Round caves Inner structure is built up on a specific flat surface, which has optimal characteristics for cell

adhesion >95% “air content“ or porosity inside the scaffold

Scaffold currently in use has been developed and patented Scaffold is made of a standard material which is a

worldwide admitted medical product (Resomer) Material combines the requested stability and the

right surface with the ability to dissolve within a defined period of time

Form: 20 mm diameter, 4 mm thickness (chip)

8

Picture of scaffold structure

9

Picture of cells on scaffold structure

10

Market and Application Area

Preclinical Development

Early Clinical Results

Content

11

Therapeutic Concept

1. First Surgery

Excision of approx. 20 to 40 gr. cirrhotic liver (from the patient) Excision of approx. 0,3 to 1,5 gr. pancreatic tissue Transport to laboratory under hypothermic conditions in PBS or

UWS (optimal time frame up to 4 hours)

2. Laboratory treatment of cells

1. Dissolving of intercellular matrix (isolation of liver cells and pancreatic islets)

Incubation of cell mix on scaffolds for approx. 30 to 60 hours Return transport to clinic (approx. 48 -65 hours upon first surgery)

§ Second Surgery Implantation of scaffolds in the peritoneum (small intestine area)

1. Follow up1. Liver function and overall condition are monitored at regular

intervalls

12

Laboratory protocol

Autologous patient samples are transported to I.A.Z.- laboratory

Principal Steps are: Perfusion of liver/ pancreas pieces with EDTA/PBS- and

then with collagenase/ hyaluronidase solution

Cutting and sieving of dissolved tissue into single cells Cells are suspended in culture medium with patients

serum Measurement of viability and number of cells Incubation of cells on polymer scaffold for 24 to 48 hours

Finally seeded scaffold chips are transported back to the clinic

13

Perfusion apparatus

14

Photographs first surgery

15

Photographs first surgery

16

Photographs second surgery

17

Photographs second surgery

18

Results of Clinical Treatment

46 Patients have received HeparAutoCell-treatment since September 2004

Patients fall into three risk categories, assessed by MELD and/or CHILD score

Suited for patients especially in the medium risk group, which enjoy higher than predicted survival time, not suited for patients in high risk group.

Relevant liver parameters have developed favorably, e.g. liver parameters have significantly improved or even normalized

4 Patients (out of the first 10) being unable to work before treatment have returned to employment

Patients listed on the transplantation list have been removed from the list due to their improved or normalized health/ liver parameters

19