Fig 2: Sampling of primary airway epithelial cells (AECs): vitality, yield, and purity. (A) Sampling method and tissue size of the conventional method or by forceps and cryo biopsy. In the conventional method, AECs were scraped off the mucosal surface after enzymatic digestion. Forceps and cryo biopsy were taken tangentially followed by enzymatic digestion. Representative pictures of tissues sizes used for cell isolation are shown for each sampling procedure. (B) Vitality directly after scraping or biopsy. Results are shown as box and whiskers (plot: min to max, show all points, n = 4). (C) Cell yield (day 8) of each sampling method. Results are shown as box and whiskers (plot: min to max, show all points, conventional: n = 11, forceps: n = 4, cryo: n = 4). (D) Flow cytometry was performed to determine cellular composition using the following markers for each cell type of AECs (CD326+): percentage of basal (CD49f+CD271+), club (CD66a/c/e+), goblet (TSPAN8+), and ciliated cells (ac. α-tubulin+). Results are shown as mean ± standard deviation. All results were analyzed by two-way analysis of variance with Tukey’s post-hoc test

Fig 3: Differentiation of airway epithelial cells (AECs) in air-liquid interface (ALI). Cellular composition of cultivated AECs on day 8, at air exposure, and after 1, 2, or 3 weeks in ALI was analyzed by flow cytometry (n = 5) using the following markers for each epithelial cell type (CD326+): (A) percentage of basal (CD49f+ CD271+), (B) club (CD66a/c/e+), (C) goblet (TSPAN8+). Results are expressed as mean ± standard deviation (plot: min to max, show all points). Statistical analyses were performed by two-way analysis of variance with Tukey’s post-hoc test