Successful retroviral gene transfer of simian virus 40 T antigen and herpes simplex virus-thymidine kinase into human hepatocytes

Current clinical reports have indicated that hepatocyte transplantation (HT X) could be used in patients with liver failure and in children with liver- based metabolic diseases. One of the major limiting factors of HTX is a ser ious shortage of donor livers for hepatocyte isolation. To address this iss ue, we immortalized adult human hepatocytes with a retroviral vector SSR#69 expressing the genes of simian virus 40 large T antigen and herpes simplex virus-thymidine kinase simultaneously. One of the resulting clones, NKNT-3 , grew steadily in chemically defined serum-free medium without any obvious crisis and showed the gene expression of differentiated liver functions. U nder the administration of 5 muM ganciclovir, NKNT-3 cells stopped prolifer ation and died in in vitro experiments. We have established a tightly regul ated immortal human hepatocyte cell line. The cells could allow the need fo r immediate availability of consistent and functionally uniform cells in su fficient quantity and adequate quality.