Primate hepatic foetal progenitor cells and their therapeutic potential

Abstract

Transplantation of genetically modified or unmodified hepatocytes appears to be a less invasive alternative to liver transplantation. However, clinical trials performed for the treatment of metabolic deficiencies resulted in a partial and transitory correction due to an insufficient number of engrafted and functional hepatocytes. In vitro, adult hepatocytes do not proliferate and the lack of organ donors limits their availability. Concomitantly, numerous works on hepatocyte transplantation in rodents have shown that cell engraftment was inefficient in normal livers. It is therefore necessary to explore the therapeutic potential of new cell sources such as stem cells and to develop pre-clinical models of transplantation. Foetal liver progenitor cells (hepatoblasts) are bipotent and express markers of both foetal hepatocytes and cholangiocytes. We have immortalized one clone of primate hepatoblasts using a retroviral vector expressing SV40 Large T and have characterized the cells at different population doublings (PDs). After 500 days in culture, immortalized cells remained bipotent and kept contact inhibition, in spite of numerous chromosomal rearrangements. After transplantation into athymic mice, the cells expressed hepatocyte functions but did not proliferate. We isolated, phenotypically characterized, transduced and cryopreserved early human hepatoblasts. These cells repopulate up to 7% of recipient immunodeficient mouse livers. This indicates that early progenitor cells display molecular characteristics related to proliferation and migration that allow these cells to engraft within hepatic parenchyma more efficiently than adult hepatocytes.