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

Jackson, Claire, Coles, Janice, Thompson, James and Lucas, Jane (2015) Seasonal variation in success and ciliation of airway epithelial cells cultured at air-liquid interface; patients samples referred for primary ciliary dyskinesia testing. European Respiratory Journal, 46 (suppl 59). (doi:10.1183/13993003.congress-2015.PA917).

Abstract

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.

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Contributors

Author: Claire Jackson

Author: James Thompson

Author: Jane Lucas

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