Experimental Lung Research, Early Online, 1–13, 2014 Copyright © 2014 Informa Healthcare USA, Inc. ISSN: 0190-2148 print / 1521-0499 online DOI: 10.3109/01902148.2014.946631 ORIGINAL ARTICLE Determinants of culture success in an airway epithelium sampling program of young children with cystic fibrosis Luke W. Garratt, 1,2 Erika N. Sutanto, 2,3 Clara J. Foo, 1 Kak Ming Ling, 2 Kevin Looi, 1 Elizabeth Kicic-Starcevich, 2,3 Thomas Iosiidis, 1,4 Kelly M. Martinovich, 2 Francis J. Lannigan, 1,5 Stephen M. Stick, 1,2,3,4 and Anthony Kicic 1,2,3,4, ∗ on behalf of AREST CF ,2,3,6,7# 1 School of Paediatrics and Child Health, University of Western Australia, Nedlands, Perth, Western Australia, Australia 2 Telethon Kids Institute, University of Western Australia, Nedlands, Perth, Western Australia, Australia 3 Department of Respiratory Medicine, Princess Margaret Hospital for Children, Perth, Western Australia, Australia 4 Centre for Cell Therapy and Regenerative Medicine, University of Western Australia, Nedlands, Perth, Western Australia, Australia 5 School of Medicine, Notre Dame University, Fremantle, Perth, Western Australia, Australia 6 Department of Respiratory Medicine, Royal Children’s Hospital, Parkville, Melbourne, Victoria, Australia 7 Murdoch Children’s Research Institute, Melbourne, Parkville, Victoria, Australia ABSTRACT Aim of the study: 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 CF ). We aimed to identify determinants of culture success through retrospective analysis of a program of routinely brushing children with CF. Materials and 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%) pAEC CF successfully cultured to passage one (P1) and 63 (24.2% of total) progressed to passage two (P2). However, >80% of non-CF specimens (pAEC non − 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 lora. Conclusions: Lower passage rates of pAEC CF cultures uniquely contrasts with pAEC non − CF despite similar cell numbers. An equivalent establishment rate of CF nasal epithelium reported elsewhere, signiicant associations to CFTR mutation phenotype, elevated airway IL-8 and opportunistic pathogens all suggest this is likely related to the CF disease milieu. KEYWORDS airway, bronchial brushing, cell culture, cystic fibrosis, epithelium Received 24 March 2014; accepted 16 July 2014 The authors would like to thank past and present fellows of the Department of Respiratory Medicine, Princess Margaret Hospital for Children, Perth, Western Australia who performed lower airway brushings. We like to thank the participants and their families who contribute to the AREST CF program. The authors also acknowledge Professor Darryl Knight for constructive criticism of the manuscript during its development. # The full membership of the Australian Respiratory Early Surveillance Team for Cystic Fibrosis (AREST CF) is available at http://www.arestcf.org Address correspondence to Anthony Kicic, Telethon Kids Institute, 100 Roberts Road, Perth, 6008 Australia. E-mail: Anthony.Kicic@telethonkids. org.au INTRODUCTION Cystic ibrosis (CF) is a fatal, inherited genetic con- dition that results from functional abnormalities of the cystic ibrosis transmembrane conductance reg- ulator (CFTR) protein, in epithelia of the lungs, gut, liver, and pancreas [1, 2]. Modern therapeu- tics have largely resolved the gastro-intestinal aspects of this multi-organ disease [3]. However lung dis- ease, featuring the structural abnormality bronchiec- tasis [4] and a susceptibility to respiratory infection, 1 Exp Lung Res Downloaded from informahealthcare.com by University of Western Australia on 09/07/14 For personal use only.