(a) Model of 3D organoid assay of airway progenitor differentiation by co-culturing sorted Gli1+ cells and ITGB4+ airway progenitors in Matrigel.
(b,c) Gli1+ mesenchyme’s effect on airway progenitor growth as demonstrated by colony forming efficiency (CFE) and organoid size of airway progenitor organoids cultured alone or with Gli1+ mesenchyme (n = 3 per group; each datapoint represents one well; one-tailed unpaired Student’s t-test). Data are expressed as mean ± SD.
(d) Cellular composition of individual airway progenitor-derived organoids.
(e-g) CFE of airway progenitors co-cultured with Gli1 Lin+ or Lin- (CD45-/CD31-/EpCAM-/YFP-) mesenchyme from PBS or bleomycin treated lung (n = 4 per group; each datapoint represents one well; one-tailed unpaired Student’s t-test). Data are expressed as mean ± SD.
(h) Proportion of airway progenitor-derived organoids with KRT5+ basal cells or SFTPC+ type 2 cells co-cultured with Gli1 Lin+ mesenchyme from bleomycin injured lungs compared to Gli1 Lin+ mesenchyme from PBS-treated lungs (n = 4 per group; each datapoint represents one well; one-tailed unpaired Student’s t-test). Data are expressed as mean ± SD. CFE = colony forming efficiency. Scale bars, 100 µm.