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Flow-Cytometric Analysis and Purification of Airway Epithelial-Cell Subsets.

American journal of respiratory cell and molecular biology (2020-11-17)
Luke R Bonser, Kyung Duk Koh, Kristina Johansson, Semil P Choksi, Dan Cheng, Leqian Liu, Dingyuan I Sun, Lorna T Zlock, Walter L Eckalbar, Walter E Finkbeiner, David J Erle
RESUMEN

The human airway epithelium is essential in homeostasis, and epithelial dysfunction contributes to chronic airway disease. Development of flow-cytometric methods to characterize subsets of airway epithelial cells will enable further dissection of airway epithelial biology. Leveraging single-cell RNA-sequencing data in combination with known cell type-specific markers, we developed panels of antibodies to characterize and isolate the major airway epithelial subsets (basal, ciliated, and secretory cells) from human bronchial epithelial-cell cultures. We also identified molecularly distinct subpopulations of secretory cells and demonstrated cell subset-specific expression of low-abundance transcripts and microRNAs that are challenging to analyze with current single-cell RNA-sequencing methods. These new tools will be valuable for analyzing and separating airway epithelial subsets and interrogating airway epithelial biology.

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Anti-CDHR3 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution