Rat tracheal epithelial cells, derived from the tracheal tissue of laboratory Sprague-Dawley rats, exhibit versatile differentiation capabilities under specific in vitro conditions. These cells can develop into mucociliary, mucous, or squamous phenotypes, with the expression of a mucociliary phenotype being most faithful to the in vivo tracheal epithelium morphology. Achieving this mucociliary phenotype necessitates the presence of retinoic acid (RA), a collagen gel substratum, and an air-liquid interface at the cellular level. In contrast, omitting RA from the culture media leads to stratified squamous cells with a cornified apical layer. This squamous phenotype is marked by the loss of transglutaminase type II and keratin 18, accompanied by the expression of squamous markers transglutaminase type I and keratin 13.
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