Establishment and characterization of an immortalized rat hepatic stellate cell line

Int J Clin Exp Pathol. 2015 Oct 1;8(10):12064-74. eCollection 2015.

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.

Keywords: Hepatic stellate cell; LSC-1; cell culture; cell line; liver fibrosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Line*
  • Hepatic Stellate Cells / cytology*
  • Hepatic Stellate Cells / metabolism
  • Immunohistochemistry
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Polymerase Chain Reaction
  • Rats
  • Tumor Protein, Translationally-Controlled 1