Immortalization of chimpanzee hepatocytes with an amphoteric retrovirus encoding simian virus 40 T antigen

Primary chimpanzee (Pan troglodytes) hepatocyte cultures were maintained in a serum-free medium containing hormones and growth factors and exhibited the de novo synthesis and secretion of numerous liver-specific plasma proteins for over 3 weeks in vitro. The long-term maintenance of differentiated, primate hepatocytes in this serum-free medium allowed for subsequent immortalization events to occur after infection with the amphoteric retrovirus U19, which encodes the simian virus 40 large T antigen oncogene. Several hepatocyte cell lines were selected and examined for the expression of liver-specific plasma proteins and the capacity to synthesize apolipoproteins. Several cell lines expressed a majority of the plasma proteins investigated, including apolipoproteins A1 and E. These results demonstrate the ability of this serum-free medium to maintain long-term differentiated primate hepatocytes, allowing for the experimental immortalization of this cell type in vitro and the maintenance of differentiated functions in the established cell lines. This methodology should be amenable to the study of the liver and its related diseases.