Fig. 5

Alveolar basal cells form organoids and differentiate to secretory- and ciliated epithelial cells. Alveolar BC were resuspended in Matrigel and cultured for 21 days as illustrated (A). Experiments presented in this figure were all performed in alveolar BC from 4 IPF patients (n-number). Representative phase contrast images of organoid formation at day 2, 7, 14, and 21 (B). Graphs, showing the number of organoids/mm³ (mean ± SEM) and the distribution of their diameters > 50 μm (violin plot) of three selected regions (1 mm³) per IPF patient (n = 4) after 20 days (C). RNA expression levels (− ΔCt) of KRT17, KRT5, TP63, SCGB1A1, MUC5AC, MUC5B, FOXJ1, or MMP7 in organoids after 21 days. Data are presented as mean ± SEM (D). Representative ICC/IF images of organoids incubated with antibodies detecting KRT17, KRT5, p63, SCGB1A1, AcTub, MUC5AC, MUC5B, or MMP7 (E), and their quantification (mean ± SEM). Dots within the bar charts represent individual data points generated by analysis of 10 organoids per IPF patient (n = 4) (F). Representative ICC/IF images of organoids incubated with antibodies detecting KRT17, KRT5, p63, SCGB1A1, AcTub, MUC5AC, MUC5B, or MMP7 (G). Nuclei were counter-stained with DAPI (G)