TY - JOUR
T1 - Establishment of immortalized human hepatic stellate scavenger cells to develop bioartificial livers
AU - Watanabe, Takamasa
AU - Shibata, Norikuni
AU - Westerman, Karen A.
AU - Okitsu, Teru
AU - Allain, Jean E.
AU - Sakaguchi, Masakiyo
AU - Totsugawa, Toshinori
AU - Maruyama, Masanobu
AU - Matsumura, Toshihisa
AU - Noguchi, Hirofumi
AU - Yamamoto, Shinichiro
AU - Hikida, Masaki
AU - Ohmori, Akira
AU - Reth, Michael
AU - Weber, Anne
AU - Tanaka, Noriaki
AU - Leboulch, Philippe
AU - Kobayashi, Naoya
PY - 2003/6/15
Y1 - 2003/6/15
N2 - Background. Maintenance of liver-specific functions has been shown to be stabilized by co-cultivation of hepatocytes with hepatic stellate cells (HSC). Because the limited lifespan of human HSC is a major hurdle to their use, the authors report here the amplification of human HSC populations in vitro by retroviral transfer of human telomerase reverse transcriptase (hTERT). Methods. Human HSC strain LI 90 cells were transduced with a retroviral vector SSR#197 expressing hTERT and green fluorescent protein (GFP) cDNA flanked by a pair of loxP. TWNT-1, one of SSR#197-immortalized HSC, was characterized. Differentiated liver functions were evaluated in an immortalized human hepatocyte NKNT-3-TWNT-1 co-culture system. Results. TWNT-1 cells showed differential functions of HSC, including uptake of acetylated low-density lipoproteins and synthesis of collagen type I and hepatocyte growth factor. Efficient excision of the retrovirally transferred hTERT and GFP cDNAs was achieved by TAT-mediated expression of the Cre recombinase and subsequent GFP-negative cell sorting. When co-cultured with TWNT-1 cells, NKNT-3 increased protein expression of the detoxifying cytochrome P450-associated protein isoenzymes 3A4 and 2C9 and urea synthesis. Conclusions. TWNT-1 cells could be valuable in the study of integrated liver functions and contribute to the optimization of liver cell therapies and bioartificial livers.
AB - Background. Maintenance of liver-specific functions has been shown to be stabilized by co-cultivation of hepatocytes with hepatic stellate cells (HSC). Because the limited lifespan of human HSC is a major hurdle to their use, the authors report here the amplification of human HSC populations in vitro by retroviral transfer of human telomerase reverse transcriptase (hTERT). Methods. Human HSC strain LI 90 cells were transduced with a retroviral vector SSR#197 expressing hTERT and green fluorescent protein (GFP) cDNA flanked by a pair of loxP. TWNT-1, one of SSR#197-immortalized HSC, was characterized. Differentiated liver functions were evaluated in an immortalized human hepatocyte NKNT-3-TWNT-1 co-culture system. Results. TWNT-1 cells showed differential functions of HSC, including uptake of acetylated low-density lipoproteins and synthesis of collagen type I and hepatocyte growth factor. Efficient excision of the retrovirally transferred hTERT and GFP cDNAs was achieved by TAT-mediated expression of the Cre recombinase and subsequent GFP-negative cell sorting. When co-cultured with TWNT-1 cells, NKNT-3 increased protein expression of the detoxifying cytochrome P450-associated protein isoenzymes 3A4 and 2C9 and urea synthesis. Conclusions. TWNT-1 cells could be valuable in the study of integrated liver functions and contribute to the optimization of liver cell therapies and bioartificial livers.
UR - http://www.scopus.com/inward/record.url?scp=0037708562&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037708562&partnerID=8YFLogxK
U2 - 10.1097/01.TP.0000064621.50907.A6
DO - 10.1097/01.TP.0000064621.50907.A6
M3 - Article
C2 - 12811248
AN - SCOPUS:0037708562
VL - 75
SP - 1873
EP - 1880
JO - Transplantation
JF - Transplantation
SN - 0041-1337
IS - 11
ER -