title: Primary outgrowth cultures are a reliable source of human pancreatic stellate cells.
abstract: Recent advances demonstrate a critical yet poorly understood role for the pancreatic stellate cell (PSC) in the pathogenesis of chronic pancreatitis (CP) and pancreatic cancer (PC). Progress in this area has been hampered by the availability, fidelity, and/or reliability of in vitro models of PSCs. We examined whether outgrowth cultures from human surgical specimens exhibited reproducible phenotypic and functional characteristics of PSCs. PSCs were cultured from surgical specimens of healthy pancreas, CP and PC. Growth dynamics, phenotypic characteristics, soluble mediator secretion profiles and co-culture with PC cells both in vitro and in vivo were assessed. Forty-seven primary cultures were established from 52 attempts, demonstrating universal α-smooth muscle actin and glial fibrillary acidic protein but negligible epithelial surface antigen expression. Modification of culture conditions consistently led to cytoplasmic lipid accumulation, suggesting induction of a quiescent phenotype. Secretion of growth factors, chemokines and cytokines did not significantly differ between donor pathologies, but did evolve over time in culture. Co-culture of PSCs with established PC cell lines resulted in significant changes in levels of multiple secreted mediators. Primary PSCs co-inoculated with PC cells in a xenograft model led to augmented tumor growth and metastasis. Therefore, regardless of donor pathology, outgrowth cultures produce PSCs that demonstrate consistent growth and protein secretion properties. Primary cultures from pancreatic surgical specimens, including malignancies, may represent a reliable source of human PSCs.
authors: Department of Surgery, College of Medicine, University of Florida Health Science Center, Gainesville, FL, USA.
authors: Song
authors: S
authors: Han
authors: Department of Surgery, College of Medicine, University of Florida Health Science Center, Gainesville, FL, USA.
authors: Daniel
authors: D
authors: Delitto
authors: Department of Surgery, College of Medicine, University of Florida Health Science Center, Gainesville, FL, USA.
authors: Dongyu
authors: D
authors: Zhang
authors: Department of Periodontology and Oral Biology, College of Dentistry, University of Florida Health Science Center, Gainesville, FL, USA.
authors: Heather L
authors: HL
authors: Sorenson
authors: Department of Surgery, College of Medicine, University of Florida Health Science Center, Gainesville, FL, USA.|Department of Surgery, North Florida/South Georgia Veterans Health System, University of Florida College of Medicine, Gainesville, FL, USA.
authors: George A
authors: GA
authors: Sarosi
authors: Department of Surgery, College of Medicine, University of Florida Health Science Center, Gainesville, FL, USA.|Department of Surgery, North Florida/South Georgia Veterans Health System, University of Florida College of Medicine, Gainesville, FL, USA.
authors: Ryan M
authors: RM
authors: Thomas
authors: Department of Surgery, College of Medicine, University of Florida Health Science Center, Gainesville, FL, USA.
authors: Kevin E
authors: KE
authors: Behrns
authors: Department of Periodontology and Oral Biology, College of Dentistry, University of Florida Health Science Center, Gainesville, FL, USA.
authors: Shannon M
authors: SM
authors: Wallet
authors: Department of Surgery, College of Medicine, University of Florida Health Science Center, Gainesville, FL, USA.
authors: Jose G
authors: JG
authors: Trevino
authors: Department of Surgery, College of Medicine, University of Florida Health Science Center, Gainesville, FL, USA.
authors: Steven J
authors: SJ
authors: Hughes
mesh_terms: D018920:Coculture Techniques; D006801:Humans; D058954:Pancreatic Stellate Cells / Q000166:cytology*; D015203:Reproducibility of Results