Skip to main content
Log in

Establishment of a human hepatocyte line (OUMS-29) having CYP 1A1 and 1A2 activities from fetal liver tissue by transfection of SV40 LT

  • Cell and Tissue Models
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Summary

Immortalized human hepatocytes that can retain functions of drug-metabolizing enzymes would be useful for medical and pharmacological studies and for constructing an artificial liver. The aim of this study was to establish immortalized human hepatocyte lines having differentiated liver-specific functions. pSVneo deoxyribonucleic acid, which contains large and small T genes in the early region of simian virus 40, was introduced into hepatocytes that had been obtained from the liver of a 21-wk-old fetus. Neomycin-resistant immortalized colonies were cloned and expanded to mass cultures to examine hepatic functions. Cells were cultured in a chemically defined serum-free medium, ASF104, which contains no peptides other than recombinant human transferrin and insulin. As a result, an immortal human hepatocyte cell line (OUMS-29) having liver-specific functions was established from one of the 13 clones. Expression of CYP 1A1 and 1A2 messenger ribonucleic acid by the cells was induced by treatment with benz[a]pyrene, 3-methylcholanthrene, and benz[a]anthracene. OUMS-29 cells had both the polycyclic aromatic hydrocarbon receptor (AhR) and AhR nuclear translocator. Consequently, 7-ethoxyresorufin deethylase activity of the cells was induced time- and dose-dependently by these polycyclic aromatic hydrocarbons. This cell line is expected to be instrumental as an alternative method in animal experiments for studying hepatocarcinogenesis, drug metabolisms of liver cells, and hepatic toxicology.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Alison, M. R. Liver stem cells: a two compartment system. Curr. Opin. Cell Biol. 10:710–715; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Alison, M. R.; Poulsom, R.; Feffery, R., et al. Hetapocytes from non-hepatic adult stem cells. Nature 406:257; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Doehmer, J.; Buters, J. T. M.; Luch, A., et al. Molecular studies on the toxifying effects by genetically engineered cytochromes P450. Drug Metab. Rev. 31:423–435; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Jacob, J. R.; Estlack, L. E.; Lanford, R. E. Immortalization of chimpanzee hepatocytes with an amphoteric retrovirus encoding simian virus 40 T antigen. Exp. Cell Res. 200:205–210; 1992.

    Article  PubMed  CAS  Google Scholar 

  • Kawada, M.; Nagamori, S.; Aizaki, H., et al. Massive culture of human liver cancer cells in a newly developed radial flow bioreactor system: ultrafine structure of functionally enhanced hepatocarcinoma cell lines. In Vitro Cell. Dev. Biol. 34A:109–115; 1998.

    Google Scholar 

  • Kono, Y.; Yang, S.; Letarte, M.; Roberts, E. A. Establishment of a human hepatocyte line derived from primary culture in a collagen gel sandwich culture system. Exp. Cell Res. 221:478–485; 1995.

    Article  PubMed  CAS  Google Scholar 

  • Lagasse, E.; Connors, H.; Al-Dhalimy, M., et al. Purified hematopoietic stem cells can differentiate into hepatocytes in vivo. Nat. Med. 6:1229–1234; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Miyazaki, M.; Mihara, K.; Bai, L., et al. Immortalization of epithelial-like cells from human liver tissue with SV40 T-antigen gene. Exp. Cell Res. 206:27–35; 1993.

    Article  PubMed  CAS  Google Scholar 

  • Miyazaki, M.; Namba, M. Hepatocellular carcinomas. In: Hay, R. J.; Park, J.-G.; Gazdar, A., ed. Atlas of human tumor cell lines. New York: Academic Press; 1994;185–212.

    Google Scholar 

  • Nawa, K.; Nakamura, T.; Kumatori, A., et al. Glucocorticoid-dependent expression of the albumin gene in adult rat hepatocytes. J. Biol. Chem. 261:16,883–16,888; 1986.

    CAS  Google Scholar 

  • Paul, D.; Hoehne, M.; Pinkert, C., et al. Immortalized differentiated hepatocyte lines derived from transgenic mice harboring SV40 T-antigen genes. Exp. Cell Res. 175:354–362; 1988.

    Article  PubMed  CAS  Google Scholar 

  • Petersen, B. E.; Bowen, W. C.; Patrene, K. D., et al. Bone marrow as a potential source of hepatic oval cells. Science 284:1168–1170; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Pfeifer, A. M. A.; Cole, K. E.; Smoot, D. T., et al. Simian virus 40 large tumor antigen-immortalized normal human liver epithelial cells express hepatocyte characteristics and metabolize chemical carcinogens. Proc. Natl. Acad. Sci. USA 90:5123–5127; 1993.

    Article  PubMed  CAS  Google Scholar 

  • Roberts, E. A.; Letarte, M.; Squire, J.; Yang, S. Characterization of human hepatocyte lines derived from normal liver tissue. Hepatology 19:1390–1399; 1994.

    Article  PubMed  CAS  Google Scholar 

  • Seglen, P.O. Preparation of rat liver cells. III. Enzymatic requirements for tissue dispersion. Exp. Cell Res. 82:391–398; 1973.

    Article  PubMed  CAS  Google Scholar 

  • Strain, A. J.; Crosby, H. A. Hepatic stem cells. Gut 46:743–745; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Theise, N. D.; Badve, S.; Saxena, R., et al. Derivation of hepatocytes from bone marrow cells in mice after radiation-induced myeloablation. Hepatology 31:235–240; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Woodworth, C. D.; Isom, H. C. Regulation of albumin gene expression in a series of rat hepatocyte cell lines immortalized by simian virus 40 and maintained in chemically defined medium. Mol. Cell. Biol. 7:3740–3748; 1987.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Masayoshi Namba.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fukaya, Ki., Asahi, S., Nagamori, S. et al. Establishment of a human hepatocyte line (OUMS-29) having CYP 1A1 and 1A2 activities from fetal liver tissue by transfection of SV40 LT. In Vitro Cell.Dev.Biol.-Animal 37, 266–269 (2001). https://doi.org/10.1007/BF02577541

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02577541

Key words

Navigation