Establishment and characterization of a simian virus 40-immortalized rat pancreatic stellate cell line

Masahiro Satoh, Atsushi Masamune, Yoshitaka Sakai, Kazuhiro Kikuta, Hirofumi Hamada, Tooru Shimosegawa

Research output: Contribution to journalReview articlepeer-review

16 Citations (Scopus)

Abstract

Activated pancreatic stellate cells (PSCs) have recently been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth potential. A continuous cell line would be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish an immortalized cell line of rat PSCs. PSCs were isolated from the pancreas of male Wistar rats, and the simian virus 40 T antigen was introduced to PSCs by retrovirus-mediated gene transfer. This procedure yielded an actively growing cell line, designated as SAM-K. This cell line has been passaged repeatedly for almost 2 years, and is thus likely immortalized. SAM-K cells retained morphological characteristics of primary PSCs, and expressed α-smooth muscle actin, glial fibrillary acidic protein, type I collagen, fibronectin, and prolyl hydroxylases. The level of p53 expression was very high in SAM-K cells. Proliferation of SAM-K cells was stimulated by serum and platelet-derived growth factor-BB. Interleukin-1β (IL-1β) activated nuclear factor-κB, activator protein-1, and three classes of mitogen-activated protein (MAP) kinases: extracellular signal-regulated kinase1/2, c-Jun N-terminal kinase, and p38 MAP kinase. IL-1β induced expression of intercellular adhesion molecule-1 and monocyte chemoattractant protein-1, both of which were abolished in the presence of pyrrolidine dithiocarbamate, a specific inhibitor of nuclear factor-κB activation. IL-1β-induced monocyte chemoattractant protein-1 was partially inhibited by specific inhibitors of MAP kinase kinase (U0126) and of p38 MAP kinase (SB203580) whereas intercellular adhesion molecule-1 expression was not altered by the inhibitors. Thus, SAM-K would be useful for in vitro studies of cell biology and signal transduction of PSCs.

Original languageEnglish
Pages (from-to)55-69
Number of pages15
JournalTohoku Journal of Experimental Medicine
Volume198
Issue number1
DOIs
Publication statusPublished - 2002 Sep 1

Keywords

  • Chronic pancreatitis
  • Pancreatic fibrosis
  • Pancreatic stellate cells
  • SV40 large T antigen
  • Signal Transduction

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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