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Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media

The culture of differentiated human airway epithelial cells allows the study of pathogen-host interactions and innate immune responses in a physiologically relevant in vitro model. As the use of primary cell culture has gained popularity the availability of the reagents needed to generate these cult...

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Autores principales: Broadbent, Lindsay, Manzoor, Sheerien, Zarcone, Maria C., Barabas, Judit, Shields, Michael D., Saglani, Sejal, Lloyd, Claire M., Bush, Andrew, Custovic, Adnan, Ghazal, Peter, Gore, Mindy, Marsland, Ben, Roberts, Graham, Schwarze, Jurgen, Turner, Steve, Power, Ultan F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7098550/
https://www.ncbi.nlm.nih.gov/pubmed/32214336
http://dx.doi.org/10.1371/journal.pone.0228229
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author Broadbent, Lindsay
Manzoor, Sheerien
Zarcone, Maria C.
Barabas, Judit
Shields, Michael D.
Saglani, Sejal
Lloyd, Claire M.
Bush, Andrew
Custovic, Adnan
Ghazal, Peter
Gore, Mindy
Marsland, Ben
Roberts, Graham
Schwarze, Jurgen
Turner, Steve
Power, Ultan F.
author_facet Broadbent, Lindsay
Manzoor, Sheerien
Zarcone, Maria C.
Barabas, Judit
Shields, Michael D.
Saglani, Sejal
Lloyd, Claire M.
Bush, Andrew
Custovic, Adnan
Ghazal, Peter
Gore, Mindy
Marsland, Ben
Roberts, Graham
Schwarze, Jurgen
Turner, Steve
Power, Ultan F.
author_sort Broadbent, Lindsay
collection PubMed
description The culture of differentiated human airway epithelial cells allows the study of pathogen-host interactions and innate immune responses in a physiologically relevant in vitro model. As the use of primary cell culture has gained popularity the availability of the reagents needed to generate these cultures has increased. In this study we assessed two different media, Promocell and PneumaCult, during the differentiation and maintenance of well-differentiated primary nasal epithelial cell cultures (WD-PNECs). We compared and contrasted the consequences of these media on WD-PNEC morphological and physiological characteristics and their responses to respiratory syncytial virus (RSV) infection. We found that cultures generated using PneumaCult resulted in greater total numbers of smaller, tightly packed, pseudostratified cells. However, cultures from both media resulted in similar proportions of ciliated and goblet cells. There were no differences in RSV growth kinetics, although more ciliated cells were infected in the PneumaCult cultures. There was also significantly more IL-29/IFNλ1 secreted from PneumaCult compared to Promocell cultures following infection. In conclusion, the type of medium used for the differentiation of primary human airway epithelial cells may impact experimental results.
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spelling pubmed-70985502020-04-03 Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media Broadbent, Lindsay Manzoor, Sheerien Zarcone, Maria C. Barabas, Judit Shields, Michael D. Saglani, Sejal Lloyd, Claire M. Bush, Andrew Custovic, Adnan Ghazal, Peter Gore, Mindy Marsland, Ben Roberts, Graham Schwarze, Jurgen Turner, Steve Power, Ultan F. PLoS One Research Article The culture of differentiated human airway epithelial cells allows the study of pathogen-host interactions and innate immune responses in a physiologically relevant in vitro model. As the use of primary cell culture has gained popularity the availability of the reagents needed to generate these cultures has increased. In this study we assessed two different media, Promocell and PneumaCult, during the differentiation and maintenance of well-differentiated primary nasal epithelial cell cultures (WD-PNECs). We compared and contrasted the consequences of these media on WD-PNEC morphological and physiological characteristics and their responses to respiratory syncytial virus (RSV) infection. We found that cultures generated using PneumaCult resulted in greater total numbers of smaller, tightly packed, pseudostratified cells. However, cultures from both media resulted in similar proportions of ciliated and goblet cells. There were no differences in RSV growth kinetics, although more ciliated cells were infected in the PneumaCult cultures. There was also significantly more IL-29/IFNλ1 secreted from PneumaCult compared to Promocell cultures following infection. In conclusion, the type of medium used for the differentiation of primary human airway epithelial cells may impact experimental results. Public Library of Science 2020-03-26 /pmc/articles/PMC7098550/ /pubmed/32214336 http://dx.doi.org/10.1371/journal.pone.0228229 Text en © 2020 Broadbent et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://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
Broadbent, Lindsay
Manzoor, Sheerien
Zarcone, Maria C.
Barabas, Judit
Shields, Michael D.
Saglani, Sejal
Lloyd, Claire M.
Bush, Andrew
Custovic, Adnan
Ghazal, Peter
Gore, Mindy
Marsland, Ben
Roberts, Graham
Schwarze, Jurgen
Turner, Steve
Power, Ultan F.
Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media
title Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media
title_full Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media
title_fullStr Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media
title_full_unstemmed Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media
title_short Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media
title_sort comparative primary paediatric nasal epithelial cell culture differentiation and rsv-induced cytopathogenesis following culture in two commercial media
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7098550/
https://www.ncbi.nlm.nih.gov/pubmed/32214336
http://dx.doi.org/10.1371/journal.pone.0228229
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