Determinants of culture success in an airway epithelium sampling program of young children with cystic fibrosis. (1st November 2014)

• Record Type: Journal Article
• Authors: Garratt, Luke W.; Sutanto, Erika N.; Foo, Clara J.; Ling, Kak Ming; Looi, Kevin; Kicic-Starcevich, Elizabeth; Iosifidis, Thomas; Martinovich, Kelly M.; Lannigan, Francis J.; Stick, Stephen M.; Kicic, Anthony; on behalf of AREST CF
• Is Part Of: Experimental lung research. Volume 40:Number 9(2014)
• Page Start: 447
• Page End: 459
• Publication Date: 2014-11-01
• Subjects: Lungs -- Periodicals; Lungs -- Diseases -- Periodicals; Lung Diseases; Lung -- physiology; Respiratory System; 616.24
• DOI: 10.3109/01902148.2014.946631
• Languages: English
• ISSNs: 0190-2148

Abstract

<abstract> <title>ABSTRACT</title> <p> <italic>Aim of the study</italic>: 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 (pAEC<sub>CF</sub>). We aimed to identify determinants of culture success through retrospective analysis of a program of routinely brushing children with CF. <italic>Materials and methods</italic>: 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. <italic>Results</italic>: Of 260 brushings processed for culture, 114 (43.8%) pAEC<sub>CF</sub> successfully cultured to passage one (P1) and 63 (24.2% of total) progressed to passage two (P2). However, >80% of non-CF specimens (pAEC<sub>non-CF</sub>) 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. <italic>Conclusions</italic>: Lower passage rates of pAEC<sub>CF</sub> cultures uniquely contrasts with pAEC<sub>non-CF</sub> 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.</p> </abstract>