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

Am J Respir Cell Mol Biol. 2021 Mar;64(3):308-317. doi: 10.1165/rcmb.2020-0149MA.


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.

Keywords: airway epithelium; flow cytometry; single-cell RNA sequencing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / metabolism
  • Biomarkers / metabolism
  • Cell Separation / methods*
  • Epithelial Cells / cytology*
  • Flow Cytometry / methods*
  • Humans
  • Respiratory System / cytology*


  • Antibodies
  • Biomarkers