Successful retroviral gene transfer of simian virus 40 T antigen and herpes simplex virus-thymidine kinase into human hepatocytes.
Current clinical reports have indicated that hepatocyte transplantation (HTX) could be used in patients with liver failure and in children with liver-based metabolic diseases. One of the major limiting factors of HTX is a serious shortage of donor livers for hepatocyte isolation. To address this issue, we immortalized adult human hepatocytes with a retroviral vector SSR#69 expressing the genes of simian virus 40 large T antigen and herpes simplex virus-thymidine kinase simultaneously. One of the resulting clones, NKNT-3, grew steadily in chemically defined serum-free medium without any obvious crisis and showed the gene expression of differentiated liver functions. Under the administration of 5 microM ganciclovir, NKNT-3 cells stopped proliferation and died in in vitro experiments. We have established a tightly regulated immortal human hepatocyte cell line. The cells could allow the need for immediate availability of consistent and functionally uniform cells in sufficient quantity and adequate quality.
Kobayashi
N
N
First Department of Surgery, Okayama University Medical School, Japan. immortal@md.okayama-u.ac.jp
Noguchi
H
H
Westerman
K A
KA
Watanabe
T
T
Matsumura
T
T
Totsugawa
T
T
Fujiwara
T
T
Leboulch
P
P
Tanaka
N
N
D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D000998:Antiviral Agents / Q000494:pharmacology; D002460:Cell Line; D017690:Cell Transplantation / Q000379:methods; D016895:Culture Media, Serum-Free; D015774:Ganciclovir / Q000494:pharmacology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology* / Q000637:transplantation; D006801:Humans; D007150:Immunohistochemistry; D008099:Liver / Q000502:physiology; D016031:Liver Transplantation; D019164:Liver, Artificial; D012190:Retroviridae / Q000235:genetics / Q000378:metabolism; D018139:Simplexvirus / Q000201:enzymology*; D013937:Thymidine Kinase / Q000235:genetics*
D000952:Antigens, Polyomavirus Transforming; D000998:Antiviral Agents; D016895:Culture Media, Serum-Free; D013937:Thymidine Kinase; D015774:Ganciclovir