Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

Abstract There is an urgent need to develop improved, physiologically-relevant in vitro models of airway epithelia with which to better understand the pathological processes associated with infection, allergies and toxicological insults of the respiratory tract of both humans and domesticated animal...

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Main Authors: Daniel Cozens, Erin Sutherland, Francesco Marchesi, Geraldine Taylor, Catherine C. Berry, Robert L. Davies
Format: Article
Language:English
Published: Nature Publishing Group 2018-10-01
Series:Scientific Reports
Subjects:
Online Access:https://doi.org/10.1038/s41598-018-33180-w
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spelling doaj-660bb38903bd4c6995c7d81d0f39ddaf2020-12-08T05:58:51ZengNature Publishing GroupScientific Reports2045-23222018-10-018111510.1038/s41598-018-33180-wTemporal differentiation of bovine airway epithelial cells grown at an air-liquid interfaceDaniel Cozens0Erin Sutherland1Francesco Marchesi2Geraldine Taylor3Catherine C. Berry4Robert L. Davies5Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of GlasgowInstitute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of GlasgowSchool of Veterinary Medicine, University of GlasgowThe Pirbright Institute, PirbrightInstitute of Molecular, Cell and Systems Biology, College of Medical, Veterinary and Life Sciences, University of GlasgowInstitute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of GlasgowAbstract There is an urgent need to develop improved, physiologically-relevant in vitro models of airway epithelia with which to better understand the pathological processes associated with infection, allergies and toxicological insults of the respiratory tract of both humans and domesticated animals. In the present study, we have characterised the proliferation and differentiation of primary bovine bronchial epithelial cells (BBECs) grown at an air-liquid interface (ALI) at three-day intervals over a period of 42 days from the introduction of the ALI. The differentiated BBEC model was highly representative of the ex vivo epithelium from which the epithelial cells were derived; a columnar, pseudostratified epithelium that was highly reflective of native airway epithelium was formed which comprised ciliated, goblet and basal cells. The hallmark defences of the respiratory tract, namely barrier function and mucociliary clearance, were present, thus demonstrating that the model is an excellent mimic of bovine respiratory epithelium. The epithelium was fully differentiated by day 21 post-ALI and, crucially, remained healthy and stable for a further 21 days. Thus, the differentiated BBEC model has a three-week window which will allow wide-ranging and long-term experiments to be performed in the fields of infection, toxicology or general airway physiology.https://doi.org/10.1038/s41598-018-33180-wNative Airway EpitheliumBovine Bronchial Epithelial Cells (BBEC)Excellent MimicPseudostratified EpitheliumTransepithelial Electrical Resistance (TEER)
collection DOAJ
language English
format Article
sources DOAJ
author Daniel Cozens
Erin Sutherland
Francesco Marchesi
Geraldine Taylor
Catherine C. Berry
Robert L. Davies
spellingShingle Daniel Cozens
Erin Sutherland
Francesco Marchesi
Geraldine Taylor
Catherine C. Berry
Robert L. Davies
Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface
Scientific Reports
Native Airway Epithelium
Bovine Bronchial Epithelial Cells (BBEC)
Excellent Mimic
Pseudostratified Epithelium
Transepithelial Electrical Resistance (TEER)
author_facet Daniel Cozens
Erin Sutherland
Francesco Marchesi
Geraldine Taylor
Catherine C. Berry
Robert L. Davies
author_sort Daniel Cozens
title Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface
title_short Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface
title_full Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface
title_fullStr Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface
title_full_unstemmed Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface
title_sort temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface
publisher Nature Publishing Group
series Scientific Reports
issn 2045-2322
publishDate 2018-10-01
description Abstract There is an urgent need to develop improved, physiologically-relevant in vitro models of airway epithelia with which to better understand the pathological processes associated with infection, allergies and toxicological insults of the respiratory tract of both humans and domesticated animals. In the present study, we have characterised the proliferation and differentiation of primary bovine bronchial epithelial cells (BBECs) grown at an air-liquid interface (ALI) at three-day intervals over a period of 42 days from the introduction of the ALI. The differentiated BBEC model was highly representative of the ex vivo epithelium from which the epithelial cells were derived; a columnar, pseudostratified epithelium that was highly reflective of native airway epithelium was formed which comprised ciliated, goblet and basal cells. The hallmark defences of the respiratory tract, namely barrier function and mucociliary clearance, were present, thus demonstrating that the model is an excellent mimic of bovine respiratory epithelium. The epithelium was fully differentiated by day 21 post-ALI and, crucially, remained healthy and stable for a further 21 days. Thus, the differentiated BBEC model has a three-week window which will allow wide-ranging and long-term experiments to be performed in the fields of infection, toxicology or general airway physiology.
topic Native Airway Epithelium
Bovine Bronchial Epithelial Cells (BBEC)
Excellent Mimic
Pseudostratified Epithelium
Transepithelial Electrical Resistance (TEER)
url https://doi.org/10.1038/s41598-018-33180-w
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