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 language | English |
|---|---|
| Pages (from-to) | 266-269 |
| Number of pages | 4 |
| Journal | In Vitro Cellular and Developmental Biology - Animal |
| Volume | 37 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2001 |
Fingerprint
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 journal › Article
}
TY - JOUR
T1 - Establishment of a human hepatocyte line (OUMS-29) having CYP 1A1 and 1A2 activities from fetal liver tissue by transfection of SV40 LT
AU - Fukaya, K. I.
AU - Asahi, S.
AU - Nagamori, S.
AU - Sakaguchi, Masakiyo
AU - Gao, C.
AU - Miyazaki, M.
AU - Namba, M.
PY - 2001
Y1 - 2001
N2 - 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.
AB - 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.
KW - 7-Ethoxyresorufin deethylase
KW - Aromatic hydrocarbon receptor
KW - Aromatic hydrocarbon receptor nuclear translocator
KW - Carcinogenic carcinogens
KW - CYP 1A1
KW - Human liver cell line
UR - http://www.scopus.com/inward/record.url?scp=0034885777&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034885777&partnerID=8YFLogxK
U2 - 10.1290/1071-2690(2001)037<0266:EOAHHL>2.0.CO;2
DO - 10.1290/1071-2690(2001)037<0266:EOAHHL>2.0.CO;2
M3 - Article
C2 - 11513080
AN - SCOPUS:0034885777
VL - 37
SP - 266
EP - 269
JO - In Vitro Cellular and Developmental Biology - Plant
JF - In Vitro Cellular and Developmental Biology - Plant
SN - 1071-2690
IS - 5
ER -