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

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

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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.

Original languageEnglish
Article numbere0228229
Number of pages12
JournalPLoS ONE
Volume15
Issue number3
DOIs
Publication statusPublished - 1 Jan 2020

Cite this

Broadbent, L., Manzoor, S., Zarcone, M. C., Barabas, J., Shields, M. D., Saglani, S., Lloyd, C. M., Bush, A., Custovic, A., Ghazal, P., Gore, M., Marsland, B., Roberts, G., Schwarze, J., Turner, S., & Power, U. F. (2020). Comparative primary paediatric nasal epithelial cell culture differentiation and RSV-induced cytopathogenesis following culture in two commercial media. PLoS ONE, 15(3), [e0228229]. https://doi.org/10.1371/journal.pone.0228229