Cre/loxP-based reversible immortalization of human hepatocytes

N. Kobayashi, H. Noguchi, K. A. Westerman, T. Watanabe, T. Matsumura, T. Totsugawa, T. Fujiwara, P. Leboulch, N. Tanaka

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22 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)383-386
Number of pages4
JournalCell Transplantation
Issue number4-5
Publication statusPublished - 2001


  • Cre/loxP system
  • Human hepatocytes
  • Reversible immortalization
  • Simian virus 40 large T antigen

ASJC Scopus subject areas

  • Biomedical Engineering
  • Cell Biology
  • Transplantation


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