TY - JOUR
T1 - Cre/loxP-based reversible immortalization of human hepatocytes
AU - Kobayashi, N.
AU - Noguchi, H.
AU - Westerman, K. A.
AU - Watanabe, T.
AU - Matsumura, T.
AU - Totsugawa, T.
AU - Fujiwara, T.
AU - Leboulch, P.
AU - Tanaka, N.
PY - 2001
Y1 - 2001
N2 - An ideal alternative to the primary human hepatocytes for hepatocyte transplantation would be to use a clonal cell line that grows economically in culture and exhibits the characteristics of differentiated, nontransformed hepatocytes following transplantation. The purpose of the present studies was to establish a reversibly immortalized human hepatocyte cell line. Human hepatocytes were immortalized with a retroviral vector SSR#69 expressing simian virus 40 large T antigen (SV40Tag) gene flanked by a pair of loxP recombination targets. One of the resulting clones, NKNT-3, showed morphological characteristics of liver parenchymal cells and expressed the genes of differentiated liver functions. NKNT-3 cells offered unlimited availability. After an adenoviral delivery of Cre recombinase and subsequent differential selection, efficient removal of SV40Tag from NKNT-3 cells was performed. Here we represent that elimination of the retrovirally transferred SV40Tag gene can be excised by adenovirus-mediated site-specific recombination.
AB - An ideal alternative to the primary human hepatocytes for hepatocyte transplantation would be to use a clonal cell line that grows economically in culture and exhibits the characteristics of differentiated, nontransformed hepatocytes following transplantation. The purpose of the present studies was to establish a reversibly immortalized human hepatocyte cell line. Human hepatocytes were immortalized with a retroviral vector SSR#69 expressing simian virus 40 large T antigen (SV40Tag) gene flanked by a pair of loxP recombination targets. One of the resulting clones, NKNT-3, showed morphological characteristics of liver parenchymal cells and expressed the genes of differentiated liver functions. NKNT-3 cells offered unlimited availability. After an adenoviral delivery of Cre recombinase and subsequent differential selection, efficient removal of SV40Tag from NKNT-3 cells was performed. Here we represent that elimination of the retrovirally transferred SV40Tag gene can be excised by adenovirus-mediated site-specific recombination.
KW - Cre/loxP system
KW - Human hepatocytes
KW - Reversible immortalization
KW - Simian virus 40 large T antigen
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U2 - 10.3727/000000001783986558
DO - 10.3727/000000001783986558
M3 - Article
C2 - 11549058
AN - SCOPUS:0034873041
VL - 10
SP - 383
EP - 386
JO - Cell Transplantation
JF - Cell Transplantation
SN - 0963-6897
IS - 4-5
ER -