Establishment of an Immortalized Human Hepatic Stellate Cell Line to Develop Antifibrotic Therapies

Cell Transplantation

Article

Masakiyo Sakaguchi¹  Noriaki Tanaka¹  Takamasa Watanabe¹  Teru Okitsu¹  Naoya Kobayashi¹  Shinichiro Yamamoto²  Kenji Omoto²  Michihiko Takesue²  Takemi Kunieda²  Norikuni Shibata² 

[1] Department of Surgery and ‡Department of Cell Biology, Okayama University Graduate School of Medicine and Dentistry, 2–5-1 Shikata-cho, Okayama 700-8558, Japan;Division of Gastroenterology, Department of Internal Medicine, Kawasaki Medical School, 577 Matsushima, Kurashiki 701-0114 Japan;

关键词: Human hepatic stellate cells;    Human telomerase reverse transcriptase;    Interferon-γ;    Rapamycin;    Type I collagen;   

DOI  :  10.3727/000000003108747064

来源: Sage Journals

【 摘 要 】

Because human hepatic stellate cells (HSCs) perform a crucial role in the progress of hepatic fibrosis, it is of great value to establish an immortalized human cell line that exhibits HSC characteristics and grows well in tissue cultures for the development of antifibrotic therapies. Thus, we engineered an immortalized human hepatic stellate cell (HSC) line TWNT-4 by retrovirally inducing human telomerase reverse transcriptase (hTERT) into LI 90 cells established from a human liver mesenchymal tumor. Parental LI 90 entered replicative senescence, whereas TWNT-4 showed telomerase activity and proliferated for more than population doubling level (PDL) 200 without any crisis. TWNT-4 expressed platelet-derived growth factor-β receptor (PDGF-βR), α-smooth muscle actin (α-SMA), and type I collagen (α1) and was considered to be an activated form of HSCs. Treatment of TWNT-4 cells with either 100 U/ml of IFN-γ or 1 ng/ml of rapamycin (Rapa) for 14 days led to lower expression of type I collagen (α1) at RNA and protein levels. Exposure of TWNT-4 cells to both of IFN-γ (10 U/ml) and Rapa (0.1 ng/ml) for 14 days effectively decreased the expression of type I collagen (α1), PDGF-βR, and α-SMA expression and suppressed TGF-β1 secretion of TWNT-4 cells. We successfully induced apoptosis by transducing TNF-related apoptosis-inducing ligand (TRAIL) into TWNT-4 cells using adenovirus vectors Ad/GT-TRAIL and Ad/PGK-GV-17. These findings suggested that immortalized activated HSC line TWNT-4 would be a useful means to develop antifibrotic therapies.

【 授权许可】

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