Seasonal variation in success and ciliation of airway epithelial cells cultured at air-liquid interface; patients samples referred for primary ciliary dyskinesia testing

Background: Air-liquid interface (ALI) culture is used to differentiate primary airway epithelial cells for cilia studies and can diagnostically confirm or excludes primary ciliary dyskinesia (PCD). This technique is capricious, time-consuming and expensive with variable success.Aims: To evaluate ALI-culture ciliation and culture failure in PCD diagnostic samples.Methods: Nasal epithelial cells from PCD referrals were cultured submerged before ALI-culture at p2 by established methods. 202 consecutive cultures (October 2013-December 2014; 27 PCD positive) were evaluated for outcome including seasonal variation.Results: 88 (43.6%; 16 PCD positive) ALI-cultures ciliated. Of 202 cultures, 72 (35.6%) were diagnostically re-analysed, 52 (25.7%) were terminated or used for research (if PCD definitely excluded by prior diagnostics) and 78 (38.6%) failed. Of 78 failed cultures: 11 failed submerged at p0 (4 non-viable, 2 fungal and 5 bacterial infections) and 67 failed at ALI (1 non-viable, 1 fungal and 2 bacterial infections, 56 unciliated and 7 no data). Seasonal variation was observed: in June-August (n=50) ciliation increased to 62.1% and culture failure decreased to 25.5%; by contrast in December-February (n=37) ciliation decreased to 25% and culture failure increased to 47.5%.Conclusion: ALI-culture complements PCD diagnostics and in vitro cilia studies. In specialist hands 44% ALI-cultures ciliate. Culture failure due to non-viable cells or bacterial or fungal infection was <5%. 28% p2 ALI-cultures failed to ciliate; winter months were suboptimal, likely associated with peaks in viral respiratory illness