title: Impact of cell culture methods on the outcomes of the in vitro inflammatory response in nasal polyps.
abstract: BACKGROUND
  In vitro culture of nasal polyp cells is frequently used in the investigation of inflammatory mechanisms and effect of treatments in nasal polyposis. Research outcomes may, however, be influenced by the culture methodology used.
  
  
  METHODS
  Nasal polyp and nasal mucosa in vitro fibroblast cultures were pre-treated with foetal bovine serum (FBS)-free culture medium or medium supplemented with either FBS or charcoal-stripped (cs) FBS. Cells were then stimulated with FBS or csFBS, with or without different doses of dexamethasone for 4 and 24h. IL-6, IL-8, GM-CSF and VEGF release and cell viability were measured.
  
  
  RESULTS
  The highest cytokine levels were found in growth-arrested cells stimulated with 10% FBS. csFBS poorly stimulated cytokine release. Nasal polyp released larger IL-8 amounts than nasal mucosa fibroblasts. Dexamethasone decreased cytokine production dose- and time-dependently in both nasal mucosa and nasal polyp fibroblasts. The IC25 of IL-8 inhibition by dexamethasone was higher in nasal polyp than in nasal mucosa fibroblasts. Cell viability did not differ among treatments.
  
  
  CONCLUSIONS
  Cytokine production by in vitro cultured nasal fibroblasts is affected by the culture conditions used and is inhibited by dexamethasone in both fibroblast types. Our results highlight the importance of culture methodology on nasal polyp research outcomes.
authors: Institut d'Investigacions Biomédiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain.
authors: Laura
authors: L
authors: Fernández-Bertolín
authors: Joaquim
authors: J
authors: Mullol
authors: Isam
authors: I
authors: Alobid
authors: Jordi
authors: J
authors: Roca-Ferrer
authors: César
authors: C
authors: Picado
authors: Laura
authors: L
authors: Pujols
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000502:physiology; D016207:Cytokines; D003907:Dexamethasone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000502:physiology; D006801:Humans; D018836:Inflammation Mediators / Q000502:physiology*; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D009297:Nasal Mucosa / Q000503:physiopathology*; D009298:Nasal Polyps / Q000503:physiopathology*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D016207:Cytokines; D018836:Inflammation Mediators; D015850:Interleukin-6; D016209:Interleukin-8; D042461:Vascular Endothelial Growth Factor A; D003907:Dexamethasone