TY - JOUR
T1 - Survival of liver failure pigs by transplantation of reversibly immortalized human hepatocytes with Tamoxifen-mediated self-recombination>
AU - Totsugawa, Toshinori
AU - Yong, Chen
AU - Rivas-Carrillo, Jorge David
AU - Soto-Gutierrez, Alejandro
AU - Navarro-Alvarez, Nalú
AU - Noguchi, Hirofumi
AU - Okitsu, Teru
AU - Westerman, Karen A.
AU - Kohara, Michinori
AU - Reth, Michael
AU - Tanaka, Noriaki
AU - Leboulch, Philippe
AU - Kobayashi, Naoya
PY - 2007/7
Y1 - 2007/7
N2 - 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.
AB - 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.
KW - Acute liver failure
KW - Hepatocyte transplantation
KW - Hepatocytes
KW - Reversible immortalization
UR - http://www.scopus.com/inward/record.url?scp=34249277092&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34249277092&partnerID=8YFLogxK
U2 - 10.1016/j.jhep.2007.02.019
DO - 10.1016/j.jhep.2007.02.019
M3 - Article
C2 - 17434229
AN - SCOPUS:34249277092
SN - 0168-8278
VL - 47
SP - 74
EP - 82
JO - Journal of Hepatology
JF - Journal of Hepatology
IS - 1
ER -