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Updated: Jul 18, 2025

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Method for Collecting Single Epithelial Cells from the Mouse Larynx.

Mohammed Imran Khan1, Meena Easwaran1,2, Joshua D Martinez1

  • 1Department of Otolaryngology - Head & Neck Surgery, School of Medicine, Stanford University, Stanford, California, U.S.A.

The Laryngoscope
|August 21, 2023
PubMed
Summary

A new dental brush method efficiently harvests pure epithelial cells from the mouse larynx. This technique enables molecular analysis of laryngeal cells after injury.

Keywords:
immunofluorescencelarynxmouse larynx epithelial cellsscRNA-seq

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Area of Science:

  • Laryngeal biology and cell isolation techniques.
  • Otorhinolaryngology research.
  • Molecular and cellular pathology.

Background:

  • The larynx contains specialized epithelial cells crucial for its function.
  • Studying molecular changes in specific laryngeal cell types requires effective isolation methods.
  • Existing methods may lead to contamination from surrounding tissues or non-epithelial cells.

Purpose of the Study:

  • To develop and validate a reliable method for harvesting individual epithelial cells from the mouse larynx.
  • To minimize contamination from non-laryngeal sites and non-epithelial laryngeal cells.
  • To enable downstream molecular and cellular assays on isolated laryngeal epithelial cells.

Main Methods:

  • A dental brush was used to collect epithelial cells from the mouse larynx after careful dissection.
  • Cells were stained for epithelial and non-epithelial markers to assess purity.
  • Histopathology evaluated the depth of tissue collection.
  • Preliminary single-cell RNA sequencing (scRNA-seq) was performed.

Main Results:

  • The brushing method yielded 6,000-8,000 cells per larynx.
  • Histopathology confirmed epithelial layer removal with minimal underlying tissue.
  • Immunofluorescence showed predominantly epithelial cell populations.
  • scRNA-seq identified nine distinct laryngeal cell clusters.

Conclusions:

  • A reliable method for harvesting pure laryngeal epithelial cells from mice was successfully developed.
  • This technique is suitable for various downstream applications, including scRNA-seq, protein analysis, and cell culture.
  • The method facilitates the study of laryngeal cells, particularly following injury.