Equine bronchial epithelial cells differentiate into ciliated and mucus producing cells in vitro.
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.
Schwab
Ute E
UE
Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA. ues3@cornell.edu
Fulcher
M Leslie
ML
Randell
Scott H
SH
Flaminio
M Julia
MJ
Russell
David G
DG
D000818:Animals; D001980:Bronchi / Q000166:cytology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D002923:Cilia / Q000648:ultrastructure; D004847:Epithelial Cells / Q000378:metabolism / Q000648:ultrastructure; D006736:Horses; D046529:Microscopy, Electron, Transmission; D055271:Mucin 5AC / Q000378:metabolism*; D055257:Mucin-5B / Q000378:metabolism*
D055271:Mucin 5AC; D055257:Mucin-5B