Journal
JOURNAL OF HEPATOLOGY
Volume 47, Issue 1, Pages 74-82Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jhep.2007.02.019
Keywords
hepatocytes; hepatocyte transplantation; acute liver failure; reversible immortalization
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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 IoxP 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. (c) 2007 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
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