title: Isolation and culture of fibroblasts from endoscopic duodenal biopsies of celiac patients.
abstract: BACKGROUND
  Fibroblasts are actually considered pivotal in inflammation and tissue remodelling process and for these reasons they are involved in the pathogenesis of autoimmune disorders such as celiac disease. Investigations to define the role of fibroblasts in celiac diseases are obstructed by the absence of specific models. Our objective is to isolate and culture primary fibroblasts from endoscopic duodenal biopsies of celiac and non-celiac subjects, to analyze their growth patterns and the morphometric characteristics.
  
  
  METHODS
  60 duodenal bioptic specimens from 20 celiac patients and 114 from 38 non-celiac subjects were mechanically chopped and enzymatically digested in order to obtain primary cell cultures. Growth patterns, karyotype (Q-banding analysis), expression of typing proteins (fibroblast surface protein and cytokeratin 20) and morphometric parameters (diameters and their ratio, perimeter, area and perimeter/area ratio at computerised image analysis) were investigated on cultured cells.
  
  
  RESULTS
  Primary cells were successfully cultured in 78% of the collected duodenal biopsies. Cultured cells, expressing the fibroblast surface protein, were negative for cytokeratine 20 and maintained a normal kariotype. Cells grew slowly without differences between the celiac and the non celiac group. Morphometric analysis of celiac fibroblasts revealed significantly increased dimensions, with a preserved diameters ratio, and a reduced perimeter/area ratio.
  
  
  CONCLUSIONS
  For the first time this study demonstrates the feasibility of culturing primary fibroblast cell from endoscopic duodenal biopsies in celiac and non-celiac subjects, opening a new window of opportunity in studies intended to establish the role of fibroblasts as a possible partaker in the pathogenesis of the celiac mucosal damage.
authors.affiliation: Center for Prevention and Diagnosis of Celiac Disease, Fondazione IRCCS Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena, Milan, Italy. leda.roncoroni@unimi.it
authors.forename: Leda
authors.initials: L
authors.lastname: Roncoroni
authors.forename: Luca
authors.initials: L
authors.lastname: Elli
authors.forename: Luisa
authors.initials: L
authors.lastname: Doneda
authors.forename: Luca
authors.initials: L
authors.lastname: Piodi
authors.forename: Michele M
authors.initials: MM
authors.lastname: Ciulla
authors.forename: Roberta
authors.initials: R
authors.lastname: Paliotti
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Bardella
mesh_terms: D000328:Adult; D001706:Biopsy; D002446:Celiac Disease / Q000473:pathology; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D004386:Duodenum / Q000473:pathology*; D004724:Endoscopy; D005260:Female; D005347:Fibroblasts / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged