Insertion of a suicide gene into an immortalized human hepatocyte cell line.
For developing a bioartificial liver (BAL) device, an attractive alternative to the primary human hepatocytes would be the use of highly differentiated immortalized human hepatocytes with a safeguard. To test the feasibility, the primary human hepatocytes were immortalized by a plasmid SV3neo encoding simian virus 40 large T antigen (SV40Tag) gene. A highly differentiated hepatocyte line OUMS-29 was established. A suicide gene of herpes simplex virus-thymidine kinase (HSV-TK) was retrovirally introduced into OUMS-29 cells as a safeguard for clinical application. One of the resulting HSV-TK-positive cell lines, OUMS-29/tk, grew in chemically defined serum-free medium with the gene expression of differentiated liver functions. OUMS-29/tk cells were 100 times more sensitive to ganciclovir compared with unmodified OUMS-29 cells in in vitro experiments. We have established a tightly regulated immortalized human hepatocyte cell line. Essentially unlimited availability of OUMS-29/tk cells may be clinically useful for BAL therapy.
Kobayashi
N
N
First Department of Surgery, Okayama University Medical School, Japan. immortal@md.okayama-u.ac.jp
Noguchi
H
H
Totsugawa
T
T
Watanabe
T
T
Matsumura
T
T
Fujiwara
T
T
Miyazaki
M
M
Fukaya
K
K
Namba
M
M
Tanaka
N
N
D000998:Antiviral Agents / Q000494:pharmacology; D002460:Cell Line; D048429:Cell Size; D016895:Culture Media, Serum-Free; D005333:Fetus; D015774:Ganciclovir / Q000494:pharmacology; D015870:Gene Expression / Q000502:physiology; D018014:Gene Transfer Techniques; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D008099:Liver; D019164:Liver, Artificial; D008297:Male; D018139:Simplexvirus / Q000201:enzymology* / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics*
D000998:Antiviral Agents; D016895:Culture Media, Serum-Free; D013937:Thymidine Kinase; D015774:Ganciclovir