RT Journal Article SR Electronic T1 Validation of an in vitro tracheal model for the study of innate host defence mechanism in respiratory tract infections JF European Respiratory Journal JO Eur Respir J FD European Respiratory Society SP P2498 VO 44 IS Suppl 58 A1 Khondoker Akram A1 Priyanka Anujan A1 Bethany Mccloskey A1 Lynne Bingle A1 Colin Bingle YR 2014 UL http://erj.ersjournals.com/content/44/Suppl_58/P2498.abstract AB In vitro models of the murine respiratory epithelium offer an attractive experimental system for discovering aspect of respiratory host defence. In contrast to the situation with human cells, murine cells have not been as extensively characterised and their host defence functions have not been fully explored. We have developed and validated an in vitro tracheal ALI (air liquid interface) culture model as a tool to understand the host defence function of the respiratory epithelium. Mouse tracheal epithelial cells (mTEC) isolated from C57BL/6 mice, were seeded onto transwells and grown to confluence in submerged culture. Cells were then cultured for 14 days at the ALI for differentiation. Transcriptional alterations were monitored during 14 days of ALI culture and apical secretions were collected for protein analysis. Our data shows that on day 14 of ALI culture the epithelial layer mimics the original tracheal epithelium. Following differentiation, SPLUNC1 and LPLUNC1, TEKT-1 (a cilia marker) and MUC5B expression was similar to that of the original cells, however CCSP expression was down-regulated compared to the original cells. SPLUNC1 protein was detected at increasing levels in the apical secretions during differentiation and the protein was localised to a non-ciliated cell population by confocal microscopy. Our study suggests that primary differentiated mTEC cultures replicate key features of the tracheal epithelium and offer an invaluable platform for subsequent studies of respiratory host defence.