Journal
EXPERIMENTAL CELL RESEARCH
Volume 330, Issue 2, Pages 300-310Publisher
ELSEVIER INC
DOI: 10.1016/j.yexcr.2014.09.033
Keywords
Human pancreatic stellate cells; Conditional immortalization; Temperature-sensitive SV4OLT; Pancreatic cancer; IGF
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Funding
- Royal Physiographic Society Lund
- Gyllenstiernska Krapperups' foundation
- John and Augusta Persson foundation
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Pancreatic stellate cells (PSCs) play a key role in the dense desmoplastic stroma associated with pancreatic ductal adenocarcinoma. Studies on human PSCs have been minimal due to difficulty in maintaining primary PSC in culture. We have generated the first conditionally immortalized human non-tumor (NPSC) and tumor-derived (TPSC) pancreatic stellate cells via transformation with the temperature-sensitive SV40 large T antigen and human telomerase (hTERT). These cells proliferate at 33 degrees C. After transfer to 37 degrees C, the SV4OLT is switched off and the cells regain their primary PSC phenotype and growth characteristics. NPSC contained cytoplasmic vitamin A-storing lipid droplets, while both NPSC and TPSC expressed the characteristic markers alpha SMA, vimentin, desmin and GFAP. Proteome array analysis revealed that of the 55 evaluated proteins, 27 (49%) were upregulated >= 3-fold in TPSC compared to NPSC, including uPA, pentraxin-3, endoglin and endothelin-1. Two insulin-like growth factor binding proteins (IGEBPs) were inversely expressed. Although discordant IGFBP-2 and IGFBP-3 levels, IGF-I was found to stimulate proliferation of both NPSC and TPSC. Both basal and IGF-I stimulated motility was significantly enhanced in TPSC compared to NPSC. In conclusion, these cells provide a unique resource that will facilitate further study of the active stroma compartment associated with pancreatic cancer. (C) 2014 The Authors. Published by Elsevier Inc.
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