Novel Method for Isolation of Murine Clara Cell Secretory Protein-Expressing Cells with Traces of Stemness

摘要: Clara cells are non-ciliated, secretory bronchiolar epithelial that serve to detoxify harmful inhaled substances. also function as stem/progenitor for repair in the bronchioles. cell protein (CCSP) is specifically expressed pulmonary and widely used a marker. In addition CCSP promoter commonly direct gene expression into lung transgenic models. The discovery of immunoreactivity plasma membranes airway lining prompted us explore possibility enriching by flow cytometry. We established novel simple method isolation CCSP-expressing using combination mechanical enzymatic dissociation followed cytometry sorting technology. showed ∼25% dissociated from whole CCSP. resulting preparation, up 98% Notably, we found several common stem markers including CD44, CD133, Sca-1 Sox2 were CCSP+ cells. Moreover, able form spheroid colonies vitro with 0.97‰ efficiency. Parallel studies vivo confirmed small population CCSP−expressing mouse airways demonstrates cell-like properties such label retention harboring rare bronchioalveolar (BASCs) terminal bronchioles (TBs). conclude exhibit number features marker expression, bronchosphere colony formation self-renewal ability. useful investigating primary research