Cell Transplantation | |
Establishment of an Immortalized Human Hepatic Stellate Cell Line to Develop Antifibrotic Therapies | |
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】
Unknown
© 2003 Cognizant Comm. Corp.
【 预 览 】
Files | Size | Format | View |
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RO202212201054432ZK.pdf | 334KB | download |
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