title: Survival of liver failure pigs by transplantation of reversibly immortalized human hepatocytes with Tamoxifen-mediated self-recombination.
abstract: OBJECTIVE
  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.
authors: Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558, Japan.
authors: Toshinori
authors: T
authors: Totsugawa
authors: Chen
authors: C
authors: Yong
authors: Jorge David
authors: JD
authors: Rivas-Carrillo
authors: Alejandro
authors: A
authors: Soto-Gutierrez
authors: Nalú
authors: N
authors: Navarro-Alvarez
authors: Hirofumi
authors: H
authors: Noguchi
authors: Teru
authors: T
authors: Okitsu
authors: Karen A
authors: KA
authors: Westerman
authors: Michinori
authors: M
authors: Kohara
authors: Michael
authors: M
authors: Reth
authors: Noriaki
authors: N
authors: Tanaka
authors: Philippe
authors: P
authors: Leboulch
authors: Naoya
authors: N
authors: Kobayashi
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002461:Cell Line, Transformed / Q000737:chemistry / Q000166:cytology / Q000637:transplantation*; D022781:Hepatocytes / Q000737:chemistry / Q000187:drug effects / Q000637:transplantation*; D006801:Humans; D019426:Integrases / Q000235:genetics; D017093:Liver Failure / Q000473:pathology / Q000601:surgery*; D011995:Recombination, Genetic; D012190:Retroviridae / Q000235:genetics; D034421:Sus scrofa; D013629:Tamoxifen / Q000494:pharmacology; D016896:Treatment Outcome
chemical_list: D015415:Biomarkers; D013629:Tamoxifen; C045073:Cre recombinase; D019426:Integrases