Temporal dynamics of ovine airway epithelial cell differentiation at an air-liquid interface

O'Boyle, N., Sutherland, E., Berry, C. C. and Davies, R. L. (2017) Temporal dynamics of ovine airway epithelial cell differentiation at an air-liquid interface. PLoS ONE, 12(7), e0181583. (doi: 10.1371/journal.pone.0181583) (PMID:28746416) (PMCID:PMC5529025)

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Abstract

The respiratory tract and lungs are subject to diverse pathologies with wide-ranging implications for both human and animal welfare. The development and detailed characterization of cell culture models for studying such forms of disease is of critical importance. In recent years the use of air-liquid interface (ALI)-cultured airway epithelial cells has increased markedly, as this method of culture results in the formation of a highly representative, organotypic in vitro model system. In this study we have expanded on previous knowledge of differentiated ovine tracheal epithelial cells by analysing the progression of differentiation over an extensive time course at an ALI. We observed a pseudo-stratified epithelium with ciliation and a concurrent increase in cell layer thickness from 9 days post-ALI with ciliation approaching a maximum level at day 24. A similar pattern was observed with respect to mucus production with intensely stained PAS-positive cells appearing at day 12. Ultrastructural analysis by SEM confirmed the presence of both ciliated cells and mucus globules on the epithelial surface within this time-frame. Trans-epithelial electrical resistance (TEER) peaked at 1049 Ω × cm2 as the cell layer became confluent, followed by a subsequent reduction as differentiation proceeded and stabilization at ~200 Ω × cm2. Importantly, little deterioration or de-differentiation was observed over the 45 day time-course indicating that the model is suitable for long-term experiments.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Berry, Dr Catherine and Davies, Dr Robert and O'Boyle, Dr Nicky and Sutherland, Miss Erin
Authors: O'Boyle, N., Sutherland, E., Berry, C. C., and Davies, R. L.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
College of Medical Veterinary and Life Sciences > Institute of Molecular Cell and Systems Biology
Journal Name:PLoS ONE
Publisher:Public Library of Science
ISSN:1932-6203
ISSN (Online):1932-6203
Copyright Holders:Copyright © 2017 O'Boyle et al.
First Published:First published in PLoS ONE 12(7): e0181583
Publisher Policy:Reproduced under a Creative Commons license

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
637741Molecular interactions of Mannheimia haemolytica with the bovine and ovine respiratory tracts using three-dimensional tissue engineering approachesRobert DaviesBiotechnology and Biological Sciences Research Council (BBSRC)BB/L010534/1III - BACTERIOLOGY