O'Boyle, N., Sutherland, E., Berry, C. C. and Davies, R. L. (2018) Optimisation of growth conditions for ovine airway epithelial cell differentiation at an air-liquid interface. PLoS ONE, 13(3), e0193998. (doi: 10.1371/journal.pone.0193998) (PMID:29518140) (PMCID:PMC5843276)
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Abstract
Respiratory tract infections are of significant concern in the agriculture industry. There is a requirement for the development of well-characterised in vitro epithelial cell culture models in order to dissect the diverse molecular interactions occurring at the host-pathogen interface in airway epithelia. We have analysed key factors that influence growth and differentiation of ovine tracheal epithelial cells in an air-liquid interface (ALI) culture system. Cellular differentiation was assessed at 21 days post-ALI, a time-point which we have previously shown to be sufficient for differentiation in standard growth conditions. We identified a dose-dependent response to epidermal growth factor (EGF) in terms of both epithelial thickening and ciliation levels. Maximal ciliation levels were observed with 25 ng ml-1 EGF. We identified a strict requirement for retinoic acid (RA) in epithelial differentiation as RA exclusion resulted in the formation of a stratified squamous epithelium, devoid of cilia. The pore-density of the growth substrate also had an influence on differentiation as high pore-density inserts yielded higher levels of ciliation and more uniform cell layers than low pore-density inserts. Differentiation was also improved by culturing the cells in an atmosphere of sub-ambient oxygen concentration. We compared two submerged growth media and observed differences in the rate of proliferation/expansion, barrier formation and also in terminal differentiation. Taken together, these results indicate important differences between the response of ovine tracheal epithelial cells and other previously described airway epithelial models, to a variety of environmental conditions. These data also indicate that the phenotype of ovine tracheal epithelial cells can be tailored in vitro by precise modulation of growth conditions, thereby yielding a customisable, potential infection model.
| 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 |
| Creator Roles: | O'Boyle, N.Data curation, Formal analysis, Investigation, Methodology, Validation, Visualization, Writing – original draft, Writing – review and editing Sutherland, E.Data curation, Investigation Berry, C. C.Project administration, Supervision, Writing – review and editing Davies, R. L.Conceptualization, Funding acquisition, Project administration, Resources, Software, Supervision, Writing – review and editing |
| Authors: | O'Boyle, N., Sutherland, E., Berry, C. C., and Davies, R. L. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity College of Medical Veterinary and Life Sciences > School of Molecular Biosciences |
| Journal Name: | PLoS ONE |
| Publisher: | Public Library of Science |
| ISSN: | 1932-6203 |
| ISSN (Online): | 1932-6203 |
| Copyright Holders: | Copyright © 2018 O'Boyle et al. |
| First Published: | First published in PLoS ONE 13(3): e0193998 |
| Publisher Policy: | Reproduced under a Creative Commons License |
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