title: Isolation and culture of primary human pancreatic stellate cells that reflect the context of their tissue of origin.
abstract: BACKGROUND
  Pancreatic stellate cells (PSCs) play a critical role in pancreatic ductal adenocarcinoma (PDAC). Activated PSCs are the main source of fibrosis in chronic pancreatitis and of desmoplasia in PDAC. The majority of studies on PSC are based on in vitro experiments relying on immortalized cell lines derived from diseased human pancreas or from animal models. These PSCs are usually activated and may not represent the biological context of their tissue of origin.
  
  
  OBJECTIVE
  (1) To isolate and culture primary human PSC from different disease contexts with minimal impact on their state of activation. (2) To perform a comparative analysis of phenotypes of PSC derived from different contexts.
  
  
  METHODS
  PSCs were isolated from normal pancreas, chronic pancreatitis, and PDAC using a hybrid method of digestion and outgrowth. To minimize activation by serum compounds, cells were cultured in a low-serum environment (2.5 % fetal bovine serum (FBS)). Expression patterns of commonly used markers for PSC phenotype and activity were compared between primary PSC lines derived from different contexts and correlated to expression in their original tissues.
  
  
  RESULTS
  Isolation was successful from 14 of 17 tissues (82 %). Isolated PSC displayed stable viability and phenotype in low-serum environment. Expression profiles of isolated PSC and matched original tissues were closely correlated. PDAC-derived PSC tended to have a higher status of activation if compared to PSC derived from non-cancerous tissues.
  
  
  CONCLUSIONS
  Primary human PSCs isolated from different contexts and cultured in a low-serum environment maintain a phenotype that reflects the stromal activity present in their tissue of origin.
authors.affiliation: Department of General Surgery, University Hospital Heidelberg, Im Neuenheimer Feld 110, 69120, Heidelberg, Germany. oliver.strobel@med.uni-heidelberg.de.
authors.forename: Oliver
authors.initials: O
authors.lastname: Strobel
authors.affiliation: Department of General Surgery, University Hospital Heidelberg, Im Neuenheimer Feld 110, 69120, Heidelberg, Germany.
authors.forename: Nigora
authors.initials: N
authors.lastname: Dadabaeva
authors.affiliation: Department of General Surgery, University Hospital Heidelberg, Im Neuenheimer Feld 110, 69120, Heidelberg, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Felix
authors.affiliation: Department of General Surgery, University Hospital Heidelberg, Im Neuenheimer Feld 110, 69120, Heidelberg, Germany.
authors.forename: Thilo
authors.initials: T
authors.lastname: Hackert
authors.affiliation: Department of General Surgery, University Hospital Heidelberg, Im Neuenheimer Feld 110, 69120, Heidelberg, Germany.
authors.forename: Nathalia A
authors.initials: NA
authors.lastname: Giese
authors.affiliation: Department of Internal Medicine 2, University Medicine Mannheim, Mannheim, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Jesenofsky
authors.affiliation: Department of General Surgery, University Hospital Heidelberg, Im Neuenheimer Feld 110, 69120, Heidelberg, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Werner
mesh_terms: D021441:Carcinoma, Pancreatic Ductal / Q000473:pathology*; D018929:Cell Culture Techniques; D002469:Cell Separation; D002470:Cell Survival; D006801:Humans; D010190:Pancreatic Neoplasms / Q000473:pathology*; D058954:Pancreatic Stellate Cells / Q000502:physiology*; D050500:Pancreatitis, Chronic / Q000473:pathology*
keywords: Activation; Chronic pancreatitis; Fibrosis; Isolation; Pancreatic cancer; Pancreatic stellate cells; Primary human