title: Establishment and characterization of an immortalized rat hepatic stellate cell line.
abstract: Hepatic stellate cells (HSCs) play an important role in liver fibrosis and portal hypertension. This study established a new rat HSC cell line LSC-1. Liver ex vivo perfusion with collagenase IV and density gradient centrifugation were used to isolate rat HSC. Cells have been maintained in culture for multiple passages. LSC-1 cell biological characteristics were studied. LSC-1 cell have been maintained in culture over 100 passages. This new HSC cell line express telomerase reverse transcriptase (TRT) and p53, suggesting that it is immortalized spontaneously. LSC-1 cells have a doubling time of 46 hours and their growth is serum-dependent. Karyotypic analysis revealed that LSC-1 cells possess normal chromosome phenotype. Moreover, LSC-1 cells do not grow in soft agar or induce tumors in nude mice, suggesting that they are not transformed. LSC-1 cells express desmin, glial fibrillary acidic proteins (GFAP), collagen type I and III, α-smooth muscle actin (α-SMA), transforming growth factor β1 (TGF-β1), platelet derived growth factor B (PDGF-B) and inducible nitric oxide synthase (iNOS). TGF-β1 stimulation increased collagen type I and III expression in LSC-1 cells. Additionally, LSC-1 cells proliferate in response to PDGF-BB, and contract in response to endothelin-1 (ET-1). In summary, LSC-1 cells exhibit activated HSC phenotype characteristics, and therefore are useful tool to study the pathogenesis of liver cirrhosis and portal hypertension.
authors.affiliation: Department of Hepatobiliary Surgery, Peking University People's Hospital Beijing, P. R. China.
authors.forename: Tao
authors.initials: T
authors.lastname: Li
authors.affiliation: Department of Hepatobiliary Surgery, Peking University People's Hospital Beijing, P. R. China.
authors.forename: Xi-Sheng
authors.initials: XS
authors.lastname: Leng
authors.affiliation: Department of Hepatobiliary Surgery, Peking University People's Hospital Beijing, P. R. China.
authors.forename: Ji-Ye
authors.initials: JY
authors.lastname: Zhu
authors.affiliation: Department of Hepatobiliary Surgery, Peking University People's Hospital Beijing, P. R. China.
authors.forename: Fu-Shun
authors.initials: FS
authors.lastname: Wang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D055166:Hepatic Stellate Cells / Q000166:cytology* / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D016133:Polymerase Chain Reaction; D051381:Rats; D000090324:Tumor Protein, Translationally-Controlled 1
keywords: Hepatic stellate cell; LSC-1; cell culture; cell line; liver fibrosis