Next Article in Journal
Oxidoreductases in Glycoprotein Glycosylation, Folding, and ERAD
Next Article in Special Issue
Ex Vivo Culture Models to Indicate Therapy Response in Head and Neck Squamous Cell Carcinoma
Previous Article in Journal
The Rationale of Neprilysin Inhibition in Prevention of Myocardial Ischemia-Reperfusion Injury during ST-Elevation Myocardial Infarction
Previous Article in Special Issue
Polysaccharide Multilayer Films in Sensors for Detecting Prostate Tumor Cells Based on Hyaluronan-CD44 Interactions
Article

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

1
Biosciences Institute, University Medical School, Newcastle University, Framlington Place, Newcastle upon Tyne NE2 4HH, UK
2
Bioinformatics Support Unit, Faculty of Medical Sciences, Newcastle University, Framlington Place, Newcastle upon Tyne NE2 4HH, UK
*
Author to whom correspondence should be addressed.
Current address: Micalis Institute, INRAE, AgroParisTech, Université Paris-Saclay, 78350 Jouy-en-Josas, France.
Cells 2020, 9(9), 2137; https://doi.org/10.3390/cells9092137
Received: 22 July 2020 / Revised: 13 September 2020 / Accepted: 16 September 2020 / Published: 21 September 2020
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. View Full-Text
Keywords: differentiation media; in vitro airway model; epithelium physiology; RNA-sequencing; pH; ion transport; cystic fibrosis; CFTR modulators differentiation media; in vitro airway model; epithelium physiology; RNA-sequencing; pH; ion transport; cystic fibrosis; CFTR modulators
Show Figures

Graphical abstract

MDPI and ACS Style

Saint-Criq, V.; Delpiano, L.; Casement, J.; Onuora, J.C.; Lin, J.; Gray, M.A. 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. Cells 2020, 9, 2137. https://doi.org/10.3390/cells9092137

AMA Style

Saint-Criq V, Delpiano L, Casement J, Onuora JC, Lin J, Gray MA. 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. Cells. 2020; 9(9):2137. https://doi.org/10.3390/cells9092137

Chicago/Turabian Style

Saint-Criq, Vinciane, Livia Delpiano, John Casement, Jennifer C. Onuora, JinHeng Lin, and Michael A. Gray. 2020. "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" Cells 9, no. 9: 2137. https://doi.org/10.3390/cells9092137

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop