Bronchiolosphere formation after co-culture of mesenchymal and club cells
(A) Whole-well overview (left), representative image of a bronchiolosphere (white square; middle) and a mesenchymal cell cluster (yellow square; right) at day 14 of co-culture of PDGFRα+ tdTom+ cells and SCGB1A1+ club cells. Scale bars represent 100 μm.
(B) Representative confocal image of an alveolosphere stained for LysoTracker (stains lamellar bodies in AECII). Scale bar represents 100 μm.
(C) Representative confocal image of a bronchiolosphere stained for SCGB1A1 (club cells) and acetylated tubulin (ciliated cells). Scale bars represent 50 μm.
(D) Whole-well overview of bronchiolospheres grown in the presence or absence of mesenchymal cells before and after passaging. Note that bronchiolospheres grown in the absence of mesenchymal cells were supplemented with rmFGF10 and rmHGF. Scale bars represent 100 μm.