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

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

1932-6203
1-12
Broadbent, Lindsay
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Manzoor, Sheerien
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Zarcone, Maria C.
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Barabas, Judit
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Shields, Michael D.
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Saglani, Sejal
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Lloyd, Claire M.
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Bush, Andrew
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Custovic, Adnan
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Ghazal, Peter
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Gore, Mindy
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Marsland, Ben
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Roberts, Graham
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Schwarze, Jurgen
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Turner, Steve
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Power, Ultan F.
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Broadbent, Lindsay
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Manzoor, Sheerien
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Zarcone, Maria C.
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Barabas, Judit
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Shields, Michael D.
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Saglani, Sejal
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Lloyd, Claire M.
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Bush, Andrew
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Custovic, Adnan
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Ghazal, Peter
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Gore, Mindy
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Marsland, Ben
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Roberts, Graham
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Schwarze, Jurgen
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Turner, Steve
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Power, Ultan F.
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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 and Power, Ultan F. (2020) Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media. PLoS ONE, 15 (3), 1-12, [e0228229]. (doi:10.1371/journal.pone.0228229).

Record type: Article

Abstract

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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Accepted/In Press date: 5 March 2020
Published date: 26 March 2020

Identifiers

Local EPrints ID: 439168
URI: http://eprints.soton.ac.uk/id/eprint/439168
ISSN: 1932-6203
PURE UUID: 8abed966-7ee1-434f-b8f7-29e303473cfd
ORCID for Graham Roberts: ORCID iD orcid.org/0000-0003-2252-1248

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Date deposited: 06 Apr 2020 16:30
Last modified: 28 Apr 2022 01:54

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Contributors

Author: Lindsay Broadbent
Author: Sheerien Manzoor
Author: Maria C. Zarcone
Author: Judit Barabas
Author: Michael D. Shields
Author: Sejal Saglani
Author: Claire M. Lloyd
Author: Andrew Bush
Author: Adnan Custovic
Author: Peter Ghazal
Author: Mindy Gore
Author: Ben Marsland
Author: Graham Roberts ORCID iD
Author: Jurgen Schwarze
Author: Steve Turner
Author: Ultan F. Power

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