Establishment and characterization of a rat pancreatic stellate cell line by spontaneous immortalization

作     者：Atsushi Masamune Masahiro Satoh Kazuhiro Kikuta Noriaki Suzuki Tooru Shimosegawa 

作者机构：Division of GastroenterologyTohoku University Graduate School of MedicineSendaiJapan A.M.and M.S.equally contributed to this work 

出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)

年 卷 期：2003年第9卷第12期

页      面：2751-2758页

核心收录：

学科分类：1002[医学-临床医学] 100201[医学-内科学(含：心血管病、血液病、呼吸系病、消化系病、内分泌与代谢病、肾病、风湿病、传染病)] 10[医学] 

基　　金：Grant-in-Aid for Encouragement of Young Scientists from Japan Society for the Promotion of Science(to A.M.) Pancreas Research Foundation of Japan(to A.M.) 

主　　题：Animals Base Sequence Cell Culture Techniques Cell Line, Transformed Cells, Cultured Cystic Fibrosis Cytoskeletal Proteins Extracellular Matrix Proteins Immunohistochemistry NF-kappa B Oligonucleotide Probes Pancreas Rats Research Support, Non-U.S. Gov't Transcription Factor AP-1 

摘      要：AIM: Activated pancreatic stellate cells (PSCs) have 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 may therefore be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish a cell line of rat PSCs by spontaneous ***: PSCs were isolated from the pancreas of male Wistar rats, and conventional subcultivation was performed repeatedly. Telomerase activity was measured using the telomere repeat amplification protocol. Activation of transcription factors was assessed by electrophoretic mobility shift *** of mitogen-activated protein (MAP) kinases was examined by Western blotting using anti-phosphospecific antibodies. Expression of cytokine-induced neutrophil chemoattractant-1 was determined by enzyme ***: Conventional subcultivation yielded actively growing cells. One clone was obtained after limiting dilution,and designated as SIPS. This cell line has been passaged repeatedly more than 2 years, and is thus likely *** cells retained morphological characteristics of primary,culture-activated PSCs. SIPS expressed α-smooth muscle actin, glial acidic fibrillary protein, vimentin, desmin, type Ⅰ collagen, fibronectin, and prolyl hydroxylases. Telomerase activity and p53 expression were negative. Proliferation of SIPS cells was serum-dependent, and stimulated with platelet-derived growth factor-BB through the activation of extracellular signal-regulated kinase. Interleukin-1β activated nuclear factor-κB, activator protein-1, and MAP ***-1β induced cytokine-induced neutrophil chemoattractant-1 expression through the activation of nuclear factor-κB and MAP ***: SIPS cells can be useful for in vitro studies of cell biology and signal transduction of PSCs.