Figure 2
Characterisation of apical-out airway organoids. (A) Number of generated organoids per well of a 24-well tissue culture plate plate across three different donors and multiple passages. Data points represent independent wells of a 24-well plate (n = 3). (B) Ciliated cell percentage of generated apical out organoids at day 15 across multiple passages. Data points represent measurements taken from independent wells of a 24-well plate (n = 3). (C) Percentage of organoids counted for panel (A) that displayed motile cilia on the outside at day 15 across three different donors and multiple passages. Organoids were classified as ‘motile’ when displaying sufficient cilia to cause motion of the organoid. (D) Histogram depicting the frequency of different Feret diameters in p3 Ap-O AO from 3 donors. 750 organoids were equally counted from 3 wells for each donor. (E) Histogram depicting the frequency of measured DAPI-positive cells in each organoid. In total 60 p3 organoids were counted. (F) Fold changes in the RNA levels of common differentiation markers in Ap-O AO across 2 different passages in three donors. Results were normalized against p3 hBECs cultured in PneumaCult-EX Plus (n = 3). (G) Expression of AC. TUB (green) and KRT5 (red) in Ap-O AO. Nuclei are visualized with DAPI (blue). Single channels are depicted in grayscale (scale bars = 50 μm). (H) Expression of ZO-1 (red) in Ap-O AO. Nuclei are visualized with DAPI (blue). Single channels are depicted in grayscale (scale bars = 50 μm).
