Isolation of alveolar type II cells and bronchioalveolar stem cells by FACS and 3D organoid culture started with FACS isolated cells carried out in a Matrigel. Panel (A): A representative plot showing the gating strategies used to isolate AT2 and BASC cells from mouse lungs. The gates were set to isolate in gate 3 AT2 (CD31 negative, CD45 negative, Ep-CAM positive, Sca-1 negative) and in gate 4 BASCs (CD31 negative, CD45 negative, Ep-CAM positive, Sca-1 positive). Panel (B): shows whole three-dimensional structure of organoid. These cultures are run for 21 days in the presence of pulmonary fibroblasts. Photograph of unstained organoid grown in insert was taken in brightfield with the use of light microscope at a 10× magnification. Panel (C): Organoid sections isolated at 21st day of culturing, paraffin embedded, 3 µm thick and H&E stained. Photograph was taken in brightfield at a 20× magnification.