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Co-culture of type I and type II pneumocytes as a model of alveolar epithelium

The epithelial tissues of the distal lung are continuously exposed to inhaled air, and are of research interest in studying respiratory exposure to both hazardous and therapeutic materials. Pharmaco-toxicological research depends on the development of sophisticated models of the alveolar epithelium,...

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Autores principales: Brookes, Oliver, Boland, Sonja, Lai Kuen, René, Miremont, Dorian, Movassat, Jamileh, Baeza-Squiban, Armelle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8475999/
https://www.ncbi.nlm.nih.gov/pubmed/34570783
http://dx.doi.org/10.1371/journal.pone.0248798
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author Brookes, Oliver
Boland, Sonja
Lai Kuen, René
Miremont, Dorian
Movassat, Jamileh
Baeza-Squiban, Armelle
author_facet Brookes, Oliver
Boland, Sonja
Lai Kuen, René
Miremont, Dorian
Movassat, Jamileh
Baeza-Squiban, Armelle
author_sort Brookes, Oliver
collection PubMed
description The epithelial tissues of the distal lung are continuously exposed to inhaled air, and are of research interest in studying respiratory exposure to both hazardous and therapeutic materials. Pharmaco-toxicological research depends on the development of sophisticated models of the alveolar epithelium, which better represent the different cell types present in the native lung and interactions between them. We developed an air-liquid interface (ALI) model of the alveolar epithelium which incorporates cell lines which bear features of type I (hAELVi) and type II (NCI-H441) epithelial cells. We compared morphology of single cells and the structure of cell layers of the two lines using light and electron microscopy. Working both in monotypic cultures and cocultures, we measured barrier function by trans-epithelial electrical resistance (TEER), and demonstrated that barrier properties can be maintained for 30 days. We created a mathematical model of TEER development over time based on these data in order to make inferences about the interactions occurring in these culture systems. We assessed expression of a panel of relevant genes that play important roles in barrier function and differentiation. The coculture model was observed to form a stable barrier akin to that seen in hAELVi, while expressing surfactant protein C, and having a profile of expression of claudins and aquaporins appropriate for the distal lung. We described cavities which arise within stratified cell layers in NCI-H441 and cocultured cells, and present evidence that these cavities represent an aberrant apical surface. In summary, our results support the coculture of these two cell lines to produce a model which better represents the breadth of functions seen in native alveolar epithelium.
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spelling pubmed-84759992021-09-28 Co-culture of type I and type II pneumocytes as a model of alveolar epithelium Brookes, Oliver Boland, Sonja Lai Kuen, René Miremont, Dorian Movassat, Jamileh Baeza-Squiban, Armelle PLoS One Research Article The epithelial tissues of the distal lung are continuously exposed to inhaled air, and are of research interest in studying respiratory exposure to both hazardous and therapeutic materials. Pharmaco-toxicological research depends on the development of sophisticated models of the alveolar epithelium, which better represent the different cell types present in the native lung and interactions between them. We developed an air-liquid interface (ALI) model of the alveolar epithelium which incorporates cell lines which bear features of type I (hAELVi) and type II (NCI-H441) epithelial cells. We compared morphology of single cells and the structure of cell layers of the two lines using light and electron microscopy. Working both in monotypic cultures and cocultures, we measured barrier function by trans-epithelial electrical resistance (TEER), and demonstrated that barrier properties can be maintained for 30 days. We created a mathematical model of TEER development over time based on these data in order to make inferences about the interactions occurring in these culture systems. We assessed expression of a panel of relevant genes that play important roles in barrier function and differentiation. The coculture model was observed to form a stable barrier akin to that seen in hAELVi, while expressing surfactant protein C, and having a profile of expression of claudins and aquaporins appropriate for the distal lung. We described cavities which arise within stratified cell layers in NCI-H441 and cocultured cells, and present evidence that these cavities represent an aberrant apical surface. In summary, our results support the coculture of these two cell lines to produce a model which better represents the breadth of functions seen in native alveolar epithelium. Public Library of Science 2021-09-27 /pmc/articles/PMC8475999/ /pubmed/34570783 http://dx.doi.org/10.1371/journal.pone.0248798 Text en © 2021 Brookes et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Brookes, Oliver
Boland, Sonja
Lai Kuen, René
Miremont, Dorian
Movassat, Jamileh
Baeza-Squiban, Armelle
Co-culture of type I and type II pneumocytes as a model of alveolar epithelium
title Co-culture of type I and type II pneumocytes as a model of alveolar epithelium
title_full Co-culture of type I and type II pneumocytes as a model of alveolar epithelium
title_fullStr Co-culture of type I and type II pneumocytes as a model of alveolar epithelium
title_full_unstemmed Co-culture of type I and type II pneumocytes as a model of alveolar epithelium
title_short Co-culture of type I and type II pneumocytes as a model of alveolar epithelium
title_sort co-culture of type i and type ii pneumocytes as a model of alveolar epithelium
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8475999/
https://www.ncbi.nlm.nih.gov/pubmed/34570783
http://dx.doi.org/10.1371/journal.pone.0248798
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