Hepatic tissue engineering: from transplantation to customized cell-based liver directed therapies from the laboratory

Abstract

Today, liver transplantation is still the only curative treatment for liver failure due to end-stages liver diseases. Donor organ shortage, high cost and the need of immunosuppressive medications are still the major limitations in the field of liver transplantation. Thus, alternative innovative cell-based liver directed therapies, e.g. liver tissue engineering, are under investigation with the aim, that in future an artificial liver tissue could be created and be used for the replacement of the liver function in patients. Using cells instead of organs in this setting should permit (i) expansion of cells in an in vitro phase, (ii) genetic or immunological manipulation of cells for transplantation, (iii) tissue typing and cryopreservation in a cell bank, and (iv) the ex vivo genetic modification of patient's own cells prior re-implantation. Function and differentiation of liver cells are influenced by the three-dimensional organ architecture. The use of polymeric matrices permits the three dimensional formation of a neo-tissue and specific stimulation by adequate modification of the matrix-surface which might be essential for appropriate differentiation of transplanted cells. Additionally, culturing hepatocytes on three dimensional matrices permits culture in a flow bioreactor system with increased function and survival of the cultured cells. Based on bioreactor technology, bioartificial liver devices (BAL) are developed for extracorporeal liver support. Although BALs improved clinical and metabolic conditions, increased patient survival rates have not been proven yet. For intra-corporeal liver replacement, a concept which combines Tissue Engineering using three-dimensional, highly porous matrices with cell transplantation could be useful. In such a concept, whole liver mass transplantation, long term engraftment and function as well as correction of a metabolic defect in animal models could be achieved with a principally reversible procedure. Future studies have to investigate, which environmental conditions and transplantation system would be most suitable for the development of artificial functional liver tissue including blood supply for a potential use in a clinical setting.

1

Scheme of the tissue engineering approach for the liver. Isolated liver cells are seeded on three-dimensional matrices in order to stimulate cell proliferation and hepatocyte-specific differentiation. The cell-seeded matrices can be used for successful three-dimensional culture, culture in a bioreactor system and for transplantation.

2

Rat hepatocytes seeded on a three-dimensional polymeric matrix form a three-dimensional tissue in vitro after 3 days in culture.

3

(A) Cell-seeded matrix at implantation between the mesenteric leaves in the peritoneal cavity. (B) Microscopic appearance of haematoxylin and eosin stained specimen of a polyvinyl-alcohol matrix seeded with freshly isolated hepatocytes. (C) H&E staining of a cell-seeded matrix 3 months after transplantation showed engrafted hepatocytes forming a neo-tissue (magnification 4 × 20).