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, Mike Shields, Sejal Saglani, Claire M. Lloyde, Andrew Bush, Adnan Custovic, Peter Ghazal, Mindy Gore, Ben Marsland, Graham Roberts, Jurgen Schwarze, Steve Turner, Ultan 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. In conclusion, the type of medium used for the differentiation of primary human airway epithelial cells impacts experimental results.
Original languageEnglish
JournalPLoS One
Early online date26 Mar 2020
DOIs
Publication statusEarly online date - 26 Mar 2020

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