Survival of liver failure pigs by transplantation of reversibly immortalized human hepatocytes with Tamoxifen-mediated self-recombination>

Toshinori Totsugawa, Chen Yong, Jorge David Rivas-Carrillo, Alejandro Soto-Gutierrez, Nalú Navarro-Alvarez, Hirofumi Noguchi, Teru Okitsu, Karen A. Westerman, Michinori Kohara, Michael Reth, Noriaki Tanaka, Philippe Leboulch, Naoya Kobayashi

Research output: Contribution to journalArticle

38 Scopus citations

Abstract

Background/Aims: Hepatocyte transplantation and bioartificial liver treatment are attractive alternatives to liver transplantation. The availability of well-characterized human hepatocyte lines facilitates such cell therapies. Methods: Human hepatocytes were immortalized with a retroviral vector SSR#197 expressing catalytic subunit of human telomerase reverse transcriptase (hTERT) and enhanced green fluorescent protein (EGFP) cDNAs flanked by a pair of loxP recombination targets. Then, Tamoxifen-dependent Cre recombinase was expressed in SSR#197-immortalized hepatocytes. Cre/LoxP recombination was performed in the established cells by simple exposure to 500 nM Tamoxifen for a week. Then, the reverted population of the cells was recovered by EGFP-negative cell sorting and characterized in vitro and in vivo using a pig model of acute liver failure (ALF) induced by d-galactosamine (0.5 g/kg) injection. Results: A human hepatocyte cell line 16T-3 was established. Reverted 16-T3 cells showed the increased expression of hepatic markers in association with enhanced levels of transcriptional factors. Compared to normal human hepatocytes, albumin production and lidocaine-metabolizing activities of reverted 16-T3 cells were 0.32 and 0.50-fold, respectively. Transplantation of reverted 16T-3 cells significantly prolonged the survival of ALF pigs. Conclusions: Here we demonstrate the usefulness of Cre/LoxP -mediated reversible immortalization of human hepatocytes with Tamoxifen-mediated self-recombination.

Original languageEnglish (US)
Pages (from-to)74-82
Number of pages9
JournalJournal of Hepatology
Volume47
Issue number1
DOIs
StatePublished - Jul 2007

Keywords

  • Acute liver failure
  • Hepatocyte transplantation
  • Hepatocytes
  • Reversible immortalization

ASJC Scopus subject areas

  • Hepatology

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    Totsugawa, T., Yong, C., Rivas-Carrillo, J. D., Soto-Gutierrez, A., Navarro-Alvarez, N., Noguchi, H., Okitsu, T., Westerman, K. A., Kohara, M., Reth, M., Tanaka, N., Leboulch, P., & Kobayashi, N. (2007). Survival of liver failure pigs by transplantation of reversibly immortalized human hepatocytes with Tamoxifen-mediated self-recombination> Journal of Hepatology, 47(1), 74-82. https://doi.org/10.1016/j.jhep.2007.02.019