Cre/loxP-Based Reversible Immortalization of Human Hepatocytes1

Cell Transplantation

Article

Toshihisa Matsumura¹  Noriaki Tanaka¹  Hirofumi Noguchi¹  Takamasa Watanabe¹  Naoya Kobayashi¹  Toshinori Totsugawa¹  Toshiyoshi Fujiwara¹  Karen A. Westerman²  Philippe Leboulch² 

[1] First Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan;Harvard-Massachusetts Institute of Technology, Division of Health Sciences and Technology, Cambridge, MA 02139;

关键词: Reversible immortalization;    Human hepatocytes;    Cre/loxP system;    Simian virus 40 large T antigen;   

DOI  :  10.3727/000000001783986558

来源: Sage Journals

【 摘 要 】

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.

【 授权许可】

Unknown   
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