Establishment of a human hepatocyte line (OUMS-29) having CYP 1A1 and 1A2 activities from fetal liver tissue by transfection of SV-10 LT

Immortalized human hepatocytes that can retain functions of drug-metabolizi ng enzymes would be useful for medical and pharmacological studies and for constructing an artificial liver. The aim of this study wa. to establish im mortalized human hepatocyte lines having differentiated liver-specific func tions. pSVneo deoxyribonucleic acid, which contains large and small T genes in the early region of simian virus 40, was introduced into hepatocytes th at had been obtained from the liver of a 21-wk-old fetus. Neomycin-resistan t immortalized colonies were cloned and expanded to mass cultures to examin e hepatic functions. Cells were cultured in a chemically defined serum-free medium, ASF101, which contains no peptides other than recombinant human tr ansferrin and insulin. As a result, air immortal human hepatocyte cell line (OUMS-29) having li er-specific functions was established from one of the 13 clones. Expression of CYP 1A1 arid 1A2 messenger ribonucleic acid by the cells was induced by treatment with benz[a]pyrene, 3-methylcholanthrene, a nd benz[a]anthracene. OUMS-29 cells had both the polycyclic aromatic hydroc arbon receptor (AhR) arid AhR nuclear translocator. Consequently, 7-ethoxyr esorufin deethylase activity of the cells was induced time- and dose-depend ent by these polycyclic aromatic hydrocarbons. This cell line is expected t o he instrumental as an alternative method in animal experiments for studyi ng hepatocarcinogenesis. drug metabolisms of liver cells, and hepatic toxic ology,.