Figure 3. Generation of optimized 2D airway organoids.

(A). A schematic graph outlines the proximal differentiation protocol to generate 2D airway organoids (AwO) in proximal differentiation medium (PD medium) at different pH. Exp medium = expansion medium. (B). 2D optimized airway organoids were applied to immunofluorescence staining to label ACCTUB+ ciliated cells (red). Nuclei and actin filaments were counterstained with DAPI (blue) and Phalloidin-647 (white), respectively. Scale bar = 20 μm.