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

K. I. Fukaya, S. Asahi, S. Nagamori, Masakiyo Sakaguchi, C. Gao, M. Miyazaki, M. Namba

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)266-269
Number of pages4
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume37
Issue number5
DOIs
Publication statusPublished - 2001

Fingerprint

Cytochrome P-450 CYP1A2
Cytochrome P-450 CYP1A1
Liver
Transfection
Hepatocytes
Fetus
Tissue
Polycyclic Aromatic Hydrocarbons
Cells
Artificial Liver
Aromatic Hydrocarbons
Cell Line
Methylcholanthrene
Simian virus 40
Neomycin
Serum-Free Culture Media
Transferrin
Cytoplasmic and Nuclear Receptors
Viruses
Metabolism

Keywords

  • 7-Ethoxyresorufin deethylase
  • Aromatic hydrocarbon receptor
  • Aromatic hydrocarbon receptor nuclear translocator
  • Carcinogenic carcinogens
  • CYP 1A1
  • Human liver cell line

ASJC Scopus subject areas

  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Establishment of a human hepatocyte line (OUMS-29) having CYP 1A1 and 1A2 activities from fetal liver tissue by transfection of SV40 LT. / Fukaya, K. I.; Asahi, S.; Nagamori, S.; Sakaguchi, Masakiyo; Gao, C.; Miyazaki, M.; Namba, M.

In: In Vitro Cellular and Developmental Biology - Animal, Vol. 37, No. 5, 2001, p. 266-269.

Research output: Contribution to journalArticle

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AU - Miyazaki, M.

AU - Namba, M.

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