Abstract
We describe a method for creating differentiated equine bronchial epithelial cell cultures that can be used for in vitro studies including airway disease mechanisms and pathogen–host interactions. Our method is based on the culturing of human tracheobronchial epithelial cells at an air–liquid interface (ALI) in specific serum-free, hormone-supplemented medium. Bronchial epithelial cells are isolated and grown on T-Clear® insert membranes. Within 2 to 3 wk, cells differentiate into ciliated and mucus producing cells as demonstrated by confocal and electron microscopy. Furthermore, the demonstration of the two major gel-forming mucin species, Muc5ac and Muc5b, in our bronchial epithelial cell culture system validates this method for studies of respiratory tract disease of the horse.
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Acknowledgements
We are grateful to Pat Burke for necropsy tissue sampling, Marlene Mardi and Shannon Caldwell for technical assistance in histology and electron microscopy, respectively.
This work was supported financially by the The Harry M. Zweig Memorial Fund for Equine Research to M. J. F. and U. E. S., and by the US Public Health Services in the form of a grant (HLD55936) from the National Heart, Lung and Blood Institute to DGH and a grant (R026-CR07) from the Cystic Fibrosis Foundation to SHR.
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Editor: J. Denry Sato
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Schwab, U.E., Fulcher, M.L., Randell, S.H. et al. Equine bronchial epithelial cells differentiate into ciliated and mucus producing cells in vitro. In Vitro Cell.Dev.Biol.-Animal 46, 102–106 (2010). https://doi.org/10.1007/s11626-009-9258-6
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DOI: https://doi.org/10.1007/s11626-009-9258-6