D.T.B. and E.S. contributed equally to this work.
Characterization of human upper airway epithelial progenitors
Article first published online: 30 JUL 2013
© 2013 ARS-AAOA, LLC
International Forum of Allergy & Rhinology
Volume 3, Issue 10, pages 841–847, October 2013
How to Cite
How to Cite this Article: Characterization of human upper airway epithelial progenitors. Int Forum Allergy Rhinol. 2013;3:841–847., , , et al.
Funding sources for the study: Stanford University Department of Otolaryngology, the Career Development Award of the Triologic Society/American Academy of Otolaryngology–Head and Neck Surgery, and the P30 Core Facilities Grant #DC010363.
Potential conflict of interest: None provided.
- Issue published online: 15 OCT 2013
- Article first published online: 30 JUL 2013
- Manuscript Accepted: 18 JUN 2013
- Manuscript Revised: 9 JUN 2013
- Manuscript Received: 15 MAR 2013
- Stanford University Department of Otolaryngology
- Career Development Award of the Triologic Society/American Academy of Otolaryngology–Head and Neck Surgery
- P30 Core Facilities. Grant Number: DC010363
- stem cells;
- nasal mucosa;
- ethmoid sinus;
- flow cytometry;
- progenitor cells;
- epithelial regeneration
New epithelial cells are generated through the proliferation and differentiation of resident progenitor cells in the nasal cavity. In several upper airway diseases, such as cystic fibrosis and chronic rhinosinusitis, self-renewing progenitor cells may be functionally defective, or compromised in their ability, to regenerate cells that maintain normal mucociliary clearance. Herein, we describe our early work to define and characterize a rare population of human nasal epithelial putative progenitors.
Single-cell suspensions of human ethmoid sinus tissues were prepared following endoscopic sinus surgery. Cell surface antibodies were analyzed as candidate markers for detecting progenitor cells. A panel of antibodies, including epithelial cell adhesion molecule (EpCAM, epithelial cells), CD45 (hematopoietic cells), nerve growth factor receptor (NGFR/CD271), intercellular adhesion molecule-1 (ICAM1/CD54), and integrin-α6 (ITGA6/CD49f) were used to resolve epithelial progenitor candidates by high-dimensional flow cytometry and the gating technique of fluorescence minus one (FMO) controls.
A rare population of approximately 0.06% of total ethmoid cells was discriminated as EpCAM−CD45−NGFR+ICAM1+ by surface markers. Use of ITGA6 was excluded based on FMO control analysis. This lineage-negative population was purified to 99% homogeneity by cell sorting and analyzed by immunofluorescence microscopy. Sorted cells were subsequently confirmed to uniformly express the transcription factor p63. Upon in vitro culture, lineage-negative clonal cells were confirmed to spontaneously differentiate into epithelial lineage-positive cells.
Using the NGFR and ICAM1 cellular coordinates, we have identified a promising population of native human nasal epithelial progenitor cells that require more formal investigation for their role in upper airway regeneration.