Background: Hepatocyte culture represents a cell model for analyzing the mechanism involved in car- cinogenesis of carcinogens, a tool for measuring hepatotoxicity of drugs and a simple model for studying hepatitis virus life cycle. However, problems raised by both the short-term survival and the poor functional stability of hepatocytes in culture hindered scientist in using of this in vitro system. Thus we planed to obtain the immortalized and differentiated human hepatocyte cell line by modifying the genome by transfecting the cells with viral specific DNA, always available in studying the liver disease. Method: After primary culture of hepatocytes obtained by therapeutic abortion at 18 weeks of gestation, SV40 large T gene was transfected into the cells by using Polybren-DMSO method, And then transfected hepatocytes were selected in G418 containing medium. Selected, transformed hepatocytes were subcultured in 10% fetal bovine serum containg F-12 medium. Morphological characteristics of subcultured cells was followed by phase-contrast microscope and electoron microscope. The immunocytochemisty using anti-human albumin and anti-human alpha fetoprotein and the immunofluorescence using snti-SV40 T antigen were performed for proving the differentiation of sub-cultured hepatocytes. Results: Electron microscope revealed subcultured cell to be epithelial cell. After more than 20 passages over a period of 7 months, the cells retained an epitheloid morphology. All the SV40 transformed hepatocyte cell lines were 100% positive for T antigen. Significant anti-alpha fetoprotein staining and week anti-albumin staining were observed in cytoplasm around nucleus and so we confirmed the synthesis of liver specific protein of transformed hepatocytes, Conclusion: Human fetal immortalized hepatocytes cell line secreting the liver specific proteins was established.