title: Determinants of culture success in an airway epithelium sampling program of young children with cystic fibrosis.
abstract: OBJECTIVE
  The bronchial brushing technique presents an opportunity to establish a gold standard in vitro model of Cystic Fibrosis (CF) airway disease. However, unique obstacles exist when establishing CF airway epithelial cells (pAECCF). We aimed to identify determinants of culture success through retrospective analysis of a program of routinely brushing children with CF.
  
  
  METHODS
  Anaesthetised children (CF and non-CF) had airway samples taken which were immediately processed for cell culture. Airway data for the CF cohort was obtained from clinical records and the AREST CF database.
  
  
  RESULTS
  Of 260 brushings processed for culture, 114 (43.8%) pAECCF successfully cultured to passage one (P1) and 63 (24.2% of total) progressed to passage two (P2). However, >80% of non-CF specimens (pAECnon-CF) cultured to P2 from similar cell numbers. Within the CF cohort, specimens successfully cultured to P2 had a higher initial cell count and lower proportion of severe CF mutation phenotype than those that did not proliferate beyond initial seeding. Elevated airway IL-8 concentration was also negatively associated with culture establishment. Contamination by opportunistic pathogens was observed in 81 (31.2% of total) cultures and brushings from children with lower respiratory tract infections were more likely to co-culture contaminating flora.
  
  
  CONCLUSIONS
  Lower passage rates of pAECCF cultures uniquely contrasts with pAECnon-CF despite similar cell numbers. An equivalent establishment rate of CF nasal epithelium reported elsewhere, significant associations to CFTR mutation phenotype, elevated airway IL-8 and opportunistic pathogens all suggest this is likely related to the CF disease milieu.
authors.affiliation: 1School of Paediatrics and Child Health, University of Western Australia, Nedlands, Perth, Western Australia, Australia.
authors.forename: Luke W
authors.initials: LW
authors.lastname: Garratt
authors.forename: Erika N
authors.initials: EN
authors.lastname: Sutanto
authors.forename: Clara J
authors.initials: CJ
authors.lastname: Foo
authors.forename: Kak Ming
authors.initials: KM
authors.lastname: Ling
authors.forename: Kevin
authors.initials: K
authors.lastname: Looi
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Kicic-Starcevich
authors.forename: Thomas
authors.initials: T
authors.lastname: Iosifidis
authors.forename: Kelly M
authors.initials: KM
authors.lastname: Martinovich
authors.forename: Francis J
authors.initials: FJ
authors.lastname: Lannigan
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Stick
authors.forename: Anthony
authors.initials: A
authors.lastname: Kicic
mesh_terms: D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry / Q000382:microbiology; D018929:Cell Culture Techniques / Q000706:statistics & numerical data*; D002648:Child; D002675:Child, Preschool; D003550:Cystic Fibrosis / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D003584:Cytological Techniques; D005260:Female; D006801:Humans; D007223:Infant; D007249:Inflammation / Q000201:enzymology; D016209:Interleukin-8 / Q000378:metabolism; D019272:Leukocyte Elastase / Q000378:metabolism; D008297:Male; D009154:Mutation; D020545:Respiratory Mucosa / Q000473:pathology*; D012189:Retrospective Studies; D013048:Specimen Handling
chemical_list: C505032:CFTR protein, human; D016209:Interleukin-8; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D019272:Leukocyte Elastase
keywords: airway; bronchial brushing; cell culture; cystic fibrosis; epithelium