Culture and Characterization of Differentiated Airway Organoids from Fetal Mouse Lung Proximal Progenitors

Developing a physiologically relevant in vitro model of the respiratory epithelium is critical for understanding lung development and respiratory diseases. Here, we describe a detailed protocol in which the fetal mouse proximal epithelial progenitors were differentiated into 3D airway organoids, which contain terminal-differentiated ciliated cells and basal stem cells. These differentiated airway organoids could constitute an excellent experimental model to elucidate the molecular mechanisms of airway development and epithelial cell fate determination and offer an important tool for establishing pulmonary dysplasia disease in vitro. Key features • Efficient isolation of proximal epithelial progenitors from mouse embryos. • Differentiation of pulmonary airway organoids differentiated from tracheal progenitors, which recapitulates the process of airway cell differentiation. • Airway organoids can be used to explore the molecular mechanisms of lung development and respiratory diseases.