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

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

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
Volume10
Issue number4-5
DOIs
Publication statusPublished - 2001

Keywords

  • 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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  • Cite this

    Kobayashi, N., Noguchi, H., Westerman, K. A., Watanabe, T., Matsumura, T., Totsugawa, T., Fujiwara, T., Leboulch, P., & Tanaka, N. (2001). Cre/loxP-based reversible immortalization of human hepatocytes. Cell Transplantation, 10(4-5), 383-386. https://doi.org/10.3727/000000001783986558