Authors:
Garratt LW, Sutanto EN, Foo CJ, Ling KM, Looi K, Kicic-Starcevich E, Iosifidis T, Martinovich KM, Lannigan FJ, Stick SM, Kicic A on behalf of AREST CF
Authors notes:
Experimental Lung Research. 2014;40(9):447-459
Keywords:
Airway, bronchial brushing, cell culture, cystic fibrosis, epithelium
Abstract:
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.
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.
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.