Figure 5.

Epithelial cell-autonomous defects in congenital lung lesions. (A) A schematic illustration of the three-dimensional lung organoid culture system. (B–F) After 2 weeks in culture, the congenital pulmonary airway malformation (CPAM) epithelium formed larger organoids, as indicated by cross-sectional area (D), with thicker walls and smaller lumens (B and C, red arrowheads). (E and G–J) H&E and immunostaining demonstrate the presence of thin-walled spherelike organoids consisting of a single cell layer of predominantly TP63⁺/KRT5⁺ basal epithelium. (F and K–N) In contrast, epithelium isolated from the CPAM lesion formed dense spheres with thick walls of TP63⁺/KRT5⁺ basal cells at least two to three cell layers thick. FACS = fluorescence-activated cell sorting; H&E = hematoxylin and eosin; KRT5 = keratin, type II cytoskeletal 5; TP63 = tumor protein 63. N = 3 patients. Four technical replicates were performed per sample. Comparison of cross-sectional area with two-tailed Student’s t test, **P < 0.01. Scale bars, 50 μm.