title: Characterization of primary cultures of cholesteatoma-associated fibroblasts.
abstract: OBJECTIVE
  The aim of this study was to characterize primary cultured fibroblasts derived from cholesteatoma (CHO) tissue to get evidence of their possible role for determining the different biological behavior of this aural pathology.
  
  
  METHODS
  Primary cultures of human fibroblasts were obtained from middle ear CHO specimens and from controls of normal human skin collected during surgical procedures. Cells were incubated with anti-vimentin monoclonal antibody, anti-cytokeratins monoclonal antibody and anti-α-smooth muscle actin monoclonal antibody. For reactive oxygen species detection, fibroblasts were incubated with 2',7'-dichlorofluorescein diacetate. Immunofluorescence, flow cytometry, confocal, and transmission electron microscopy were carried out.
  
  
  RESULTS
  Cholesteatoma-associated fibroblasts (CHO-AFs) were characterized by a higher degree of cytoplasmic complexity, by the expression of α-smooth muscle actin and by a greater basal production of reactive oxygen species in comparison with controls, reflecting a more differentiated phenotype consistent with myofibroblasts.
  
  
  CONCLUSIONS
  It is possible to suggest that the differentiated phenotype of CHO-AFs might be coupled with a more or less aggressive clinical behavior, and hence, these cultures might represent powerful tools for investigating biology and clinical evolution of this disease.
authors: Department NESMOS, Otorhinolaryngology, University La Sapienza, Rome, Italy. maurizio.barbara@uniroma1.it
authors: Maurizio
authors: M
authors: Barbara
authors: Salvatore
authors: S
authors: Raffa
authors: Laura
authors: L
authors: Leone
authors: Carmelo
authors: C
authors: Murè
authors: Cristina
authors: C
authors: Scrofani
authors: Simonetta
authors: S
authors: Monini
authors: Maria Rosaria
authors: MR
authors: Torrisi
mesh_terms: D000199:Actins / Q000378:metabolism; D002478:Cells, Cultured; D002781:Cholesteatoma / Q000473:pathology*; D004424:Ear Canal / Q000002:abnormalities / Q000473:pathology; D005347:Fibroblasts / Q000473:pathology* / Q000648:ultrastructure; D005434:Flow Cytometry; D005452:Fluoresceins; D005455:Fluorescent Antibody Technique; D006801:Humans; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D010641:Phenotype; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D005452:Fluoresceins; D017382:Reactive Oxygen Species; C037631:2',7'-dichlorofluorescein