Establishment and Characterization of a Rat Pancreatic Stellate Cell Line by Spontaneous Immortalization

Overview

Journal World J Gastroenterol

Publisher Baishideng Publishing Group

Specialty Gastroenterology

Date 2003 Dec 12

PMID 14669327

Citations 14

Authors

Atsushi Masamune

Masahiro Satoh

Kazuhiro Kikuta

Noriaki Suzuki

Tooru Shimosegawa

Affiliations

Soon will be listed here.

Abstract

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 immortalization.

Methods: 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 assay. Activation 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 immunoassay.

Results: 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 immortalized. SIPS cells retained morphological characteristics of primary, culture-activated PSCs. SIPS expressed alpha-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-1beta activated nuclear factor-kappaB, activator protein-1, and MAP kinases. Interleukin-1beta induced cytokine-induced neutrophil chemoattractant-1 expression through the activation of nuclear factor-kappaB and MAP kinases.

Conclusion: SIPS cells can be useful for in vitro studies of cell biology and signal transduction of PSCs.

Citing Articles

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Che M, Kweon S, Teo J, Yuan Y, Melstrom L, Waldron R Cancers (Basel). 2020; 12(6).

PMID: 32516943 PMC: 7352534. DOI: 10.3390/cancers12061476.

Feeder-cell-independent culture of the pig embryonic stem cell-derived exocrine pancreatic cell line, PICM-31.

Talbot N, Shannon A, Phillips C, Garrett W In Vitro Cell Dev Biol Anim. 2018; 54(4):321-330.

PMID: 29442225 DOI: 10.1007/s11626-017-0218-2.

Pancreatic stellate cell: Pandora's box for pancreatic disease biology.

Bynigeri R, Jakkampudi A, Jangala R, Subramanyam C, Sasikala M, Venkat Rao G World J Gastroenterol. 2017; 23(3):382-405.

PMID: 28210075 PMC: 5291844. DOI: 10.3748/wjg.v23.i3.382.

Characterization of Long-Term Cultured Murine Submandibular Gland Epithelial Cells.

Ikeura K, Kawakita T, Tsunoda K, Nakagawa T, Tsubota K PLoS One. 2016; 11(1):e0147407.

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Spontaneous immortalization of mouse liver sinusoidal endothelial cells.

Zhao X, Zhao Q, Luo Z, Yu Y, Xiao N, Sun X Int J Mol Med. 2015; 35(3):617-24.

PMID: 25585915 PMC: 4314414. DOI: 10.3892/ijmm.2015.2067.

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