title: Growth and differentiation of primary and passaged equine bronchial epithelial cells under conventional and air-liquid-interface culture conditions.
abstract: BACKGROUND
  Horses develop recurrent airway obstruction (RAO) that resembles human bronchial asthma. Differentiated primary equine bronchial epithelial cells (EBEC) in culture that closely mimic the airway cells in vivo would be useful to investigate the contribution of bronchial epithelium in inflammation of airway diseases. However, because isolation and characterization of EBEC cultures has been limited, we modified and optimized techniques of generating and culturing EBECs from healthy horses to mimic in vivo conditions.
  
  
  RESULTS
  Large numbers of EBEC were obtained by trypsin digestion and successfully grown for up to 2 passages with or without serum. However, serum or ultroser G proved to be essential for EBEC differentiation on membrane inserts at ALI. A pseudo-stratified muco-ciliary epithelium with basal cells was observed at differentiation. Further, transepithelial resistance (TEER) was more consistent and higher in P1 cultures compared to P0 cultures while ciliation was delayed in P1 cultures.
  
  
  CONCLUSIONS
  This study provides an efficient method for obtaining a high-yield of EBECs and for generating highly differentiated cultures. These EBEC cultures can be used to study the formation of tight junction or to identify epithelial-derived inflammatory factors that contribute to lung diseases such as asthma.
authors.affiliation: Institute of Pharmacology, Pharmacy and Toxicology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 15, 04103 Leipzig, Germany. gabraham@rz.uni-leipzig.de
authors.forename: Getu
authors.initials: G
authors.lastname: Abraham
authors.forename: Claudia
authors.initials: C
authors.lastname: Zizzadoro
authors.forename: Johannes
authors.initials: J
authors.lastname: Kacza
authors.forename: Christin
authors.initials: C
authors.lastname: Ellenberger
authors.forename: Vanessa
authors.initials: V
authors.lastname: Abs
authors.forename: Jana
authors.initials: J
authors.lastname: Franke
authors.forename: Heinz-Adolf
authors.initials: HA
authors.lastname: Schoon
authors.forename: Johannes
authors.initials: J
authors.lastname: Seeger
authors.forename: Yohannes
authors.initials: Y
authors.lastname: Tesfaigzi
authors.forename: Fritz R
authors.initials: FR
authors.lastname: Ungemach
mesh_terms: D000818:Animals; D001980:Bronchi / Q000166:cytology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002999:Clone Cells; D003470:Culture Media; D016895:Culture Media, Serum-Free; D006736:Horses; D020545:Respiratory Mucosa / Q000166:cytology*
chemical_list: D003470:Culture Media; D016895:Culture Media, Serum-Free