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Choice of Differentiation Media Significantly Impacts Cell Lineage and Response to CFTR Modulators in Fully Differentiated Primary Cultures of Cystic Fibrosis Human Airway Epithelial Cells

Lookup NU author(s): Dr Vinciane Saint-CriqORCiD, Livia Delpiano, John CasementORCiD, Jennifer Chika Onuora, Jia-Yu Lin, Dr Michael Gray



This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


In vitro cultures of primary human airway epithelial cells (hAECs) grown at air-liquid interface have become a valuable tool to study airway biology under normal and pathologic conditions, and for drug discovery in lung diseases such as cystic fibrosis (CF). An increasing number of different differentiation media, are now available, making comparison of data between studies difficult. Here, we investigated the impact of two common differentiation media on phenotypic, transcriptomic, and physiological features of CF and non-CF epithelia. Cellular architecture and density were strongly impacted by the choice of medium. RNA-sequencing revealed a shift in airway cell lineage; one medium promoting differentiation into club and goblet cells whilst the other enriched the growth of ionocytes and multiciliated cells. Pathway analysis identified differential expression of genes involved in ion and fluid transport. Physiological assays (intracellular/extracellular pH, Ussing chamber) specifically showed that ATP12A and CFTR function were altered, impacting pH and transepithelial ion transport in CF hAECs. Importantly, the two media differentially affected functional responses to CFTR modulators. We argue that the effect of growth conditions should be appropriately determined depending on the scientific question and that our study can act as a guide for choosing the optimal growth medium for specific applications.

Publication metadata

Author(s): Saint-Criq V, Delpiano L, Casement J, Onuora JC, Lin J, Gray MA

Publication type: Article

Publication status: Published

Journal: Cells

Year: 2020

Volume: 9

Issue: 9

Online publication date: 21/09/2020

Acceptance date: 16/09/2020

Date deposited: 10/12/2020

ISSN (electronic): 2073-4409

Publisher: MDPI


DOI: 10.3390/cells9092137

PubMed id: 32967385


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