Pharmacological Rescue of Conventional Primary Equine Bronchial Epithelial Cells

Abstract

Aim: Well-differentiated primary equine bronchial epithelial (EBEC) cell cultures are pivotal for airway disease research, particularly for development of drugs to treat equine recurrent airway obstruction (RAO) and human asthma. Horses often suffer from an asthma-like airway disease and can be used as large animal models. We initially determined the electrophysiological and morphological characteristics of EBEC, cultured under conventional and air liquid interface (ALI) conditions and here, the influence of the RhoA kinase (ROCK) inhibitor Y-27632 (Y). Material and Methods: Fresh isolated EBECs were cultured in the presence and absence of Y-27632 under conventional and ALI conditions in supplemented airway epithelial cell growth medium. The effects of Y-27632 on cell viability, morphology, proliferation and differentiation were examined. Results: When treating EBEC cultures with the inhibitor, cells grew exponentially for a number of passages (>4) with less contamination of the EBEC with fibroblasts. Passaged EBEC formed confluent mucociliary ALI cultures. There were differences in cell morphology, tight junction formation and current magnitude as a function of extended passages between Y-27632-treated and not treated EBEC. Conclusion: The ability to rapidly generate many primary EBE cells from bronchial specimens provides significant opportunities for cell-based diagnostics and therapeutics. This study confirms the benefits of Y-27632 to provide the opportunity to culture primary EBEC for extended periods of culture.