title: Conditionally immortalized human pancreatic stellate cell lines demonstrate enhanced proliferation and migration in response to IGF-I.
abstract: 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°C. After transfer to 37°C, the SV40LT 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 αSMA, 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 (IGFBPs) 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.
authors.affiliation: Lund University, Department of Clinical Sciences Lund, Division of Surgery, Lund, Sweden; Lund University and Skåne University Hospital, Department of Clinical Sciences Lund, Division of Oncology and Pathology, Lund, Sweden. Electronic address: ann.rosendahl@med.lu.se.
authors.forename: Ann H
authors.initials: AH
authors.lastname: Rosendahl
authors.affiliation: Lund University, Department of Clinical Sciences Lund, Division of Surgery, Lund, Sweden.
authors.forename: Chinmay
authors.initials: C
authors.lastname: Gundewar
authors.affiliation: Lund University, Department of Clinical Sciences Lund, Division of Surgery, Lund, Sweden.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Said Hilmersson
authors.affiliation: University of Bristol, School of Clinical Sciences, Children׳s Renal Unit and Academic Renal Unit, Bristol, UK.
authors.forename: Lan
authors.initials: L
authors.lastname: Ni
authors.affiliation: University of Bristol, School of Clinical Sciences, Children׳s Renal Unit and Academic Renal Unit, Bristol, UK.
authors.forename: Moin A
authors.initials: MA
authors.lastname: Saleem
authors.affiliation: Lund University, Department of Clinical Sciences Lund, Division of Surgery, Lund, Sweden.
authors.forename: Roland
authors.initials: R
authors.lastname: Andersson
mesh_terms: D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D021441:Carcinoma, Pancreatic Ductal / Q000473:pathology*; D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000502:physiology; D002465:Cell Movement; D049109:Cell Proliferation; D003893:Desmin / Q000096:biosynthesis; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis; D006801:Humans; D018971:Insulin-Like Growth Factor Binding Protein 2 / Q000378:metabolism; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000378:metabolism / Q000494:pharmacology*; D009361:Neoplasm Invasiveness / Q000473:pathology; D010183:Pancreatic Ducts / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000473:pathology*; D058954:Pancreatic Stellate Cells / Q000378:metabolism*; D061251:Primary Cell Culture; D051785:Smad Proteins / Q000096:biosynthesis; D019098:Telomerase / Q000235:genetics; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000096:biosynthesis
chemical_list: D000952:Antigens, Polyomavirus Transforming; D003893:Desmin; D005904:Glial Fibrillary Acidic Protein; C577903:IGFBP2 protein, human; C515497:IGFBP3 protein, human; D018971:Insulin-Like Growth Factor Binding Protein 2; D018972:Insulin-Like Growth Factor Binding Protein 3; D051785:Smad Proteins; D014746:Vimentin; D007334:Insulin-Like Growth Factor I; D019098:Telomerase
keywords: Conditional immortalization; Human pancreatic stellate cells; IGF; Pancreatic cancer; Temperature-sensitive SV40LT