Dhivya Haridass

Hepatocyte transplantation has been proposed as an attractive therapeutic approach to a variety of liver diseases. Progress, however, in several areas is needed, before this form of therapy is broadly accepted and applied to patients with liver disease. The purpose of the review is to provide the reader with the latest scientific developments relevant for future liver cell therapies. Clinical application of hepatocyte transplantation is limited by good quality donor livers for the isolation of cells. Recent publications thus focus on stem cells as suitable sources for hepatocytes and liver repopulation strategies that could reduce the number of transplanted cells. How to overcome host immune responses against allogeneic cells can potentially be learned from a new tolerance protocol. Recently discovered technologies for reprogramming of postnatal cells into pluripotent stem cells may pave the way towards the generation of patient-specific autologous cells. The current focus of research aims to reduce the shortage of transplantable cells by the application of stem cell sources or by the conditioning of recipient livers. Therapies for severe chronic liver diseases based on adult (stem) cells are already beginning to move into clinical trials. However, many questions about safety and efficacy need to be answered, before fetal liver progenitor cells, embryonic stem cells and induced pluripotent stem cells can be applied in humans.

Directed endodermal differentiation of murine embryonic stem (ES) cells gives rise to a subset of cells with a hepatic phenotype. Such ES cell-derived hepatic progenitor cells (ES-HPC) can acquire features of hepatocytes in vitro, but fail to form substantial hepatocyte clusters in vivo. In this study, we investigated whether this is due to inefficient engraftment or an immature phenotype of ES-HPC. ES cells engrafted into recipient livers of NOD/SCID mice with a similar efficacy as adult hepatocytes after 28 days. Because transplanted unpurified ES-HPC formed teratomas in the spleen and liver, we applied an albumin promoter/enhancer-driven reporter system to purify ES-HPC by cell sorting. RT-PCR analyses for hepatocyte-specific genes showed that the cells exhibited a hepatic phenotype, lacking the expression of the pluripotency marker Oct4, comparable to cells of day 11.5 embryos. Sorted ES-HPC derived from β-galactosidase transgenic ES cells were injected into fumaryl-acetoacetate-deficient (FAH-/-) SCID mice and analyzed after 8 to 12 weeks. Staining with X-gal solution revealed the presence of engrafted cells throughout the liver. However, immunostaining for the FAH protein indicated hepatocyte formation at a very low frequency, without evidence for large hepatocyte cluster formation. In conclusion, the limited repopulation capacity of ES-HPC is not caused by a failure of primary engraftment, but may be due to an immature hepatic phenotype of the transplanted ES-HPC.