Culture and differentiation of mouse tracheal epithelial cells

Methods Mol Biol. 2013;945:123-43. doi: 10.1007/978-1-62703-125-7_9.


Airway epithelial cell biology has been greatly advanced by studies of genetically defined and modified mice; however it is often difficult to isolate, manipulate, and assay epithelial cell-specific responses in vivo. In vitro proliferation and differentiation of mouse airway epithelial cells are made possible by a high-fidelity system for primary culture of mouse tracheal epithelial cells described in this chapter. Using this method, epithelial cells purified from mouse tracheas proliferate in growth factor-enriched medium. Subsequent culture in defined medium and the use of the air-liquid interface condition result in the development of well-differentiated epithelia composed of ciliated and non-ciliated cells with characteristics of native airways. Methods are also provided for manipulation of differentiation and analysis of differentiation and gene expression. These approaches allow the assessment of global responses and those of specific cell subpopulations within the airway epithelium.

Publication types

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

MeSH terms

  • Air
  • Animals
  • Cell Culture Techniques / methods*
  • Cell Differentiation*
  • Cell Proliferation
  • Cell Separation
  • Cryopreservation
  • Culture Media / chemistry
  • Epithelial Cells / cytology*
  • Epithelial Cells / metabolism
  • Fluorescent Antibody Technique
  • Gene Expression Profiling
  • Mice
  • Trachea / cytology*


  • Culture Media