Figure 3
Development of non-celiac and celiac organoid-derived monolayers. (a) Transepithelial electrical resistance (TEER) measurement to evaluate development of the monolayers over time. Data represent average ± SEM. **p < 0.01, ***p < 0.001, two-side unpaired test; 149 to 234 replicates for n = 5 non-celiac (NC) and 156 to 253 replicates for n = 5 active celiac (CD) organoid-derived monolayers. (b,c) Absolute fluorescence intensity of FITC-dextran of 4,000 Da (a) and FITC-PEG of 400 Da (b) recovered in the basolateral side after 4 h of incubation. Data represent average ± SEM. (b) *p < 0.05, two-side unpaired t-test; 12 replicates for n = 4 non-celiac (NC) and n = 4 active celiac (CD) at each time point. (c) **p < 0.01, ***p < 0.001, two-side unpaired t-test; 12 and 9 replicates for n = 4 non-celiac (NC) and n = 3 active celiac (CD) at each time point. (d) Immunofluorescence staining of TJP1 (also known as ZO1) performed on one representative non-celiac (NC) and one active celiac (CD) monolayer at day 7 showing reduced deposition of the tight junction protein along the apical cell perimeter. (e–h) Gene expression assessed by qRT-PCR in organoid-derived monolayers cultured in two differentiation conditions (see Methods). LYZ (e), SI (f), MUC2 (g) and CHGA (h) evaluated the relative abundance of Paneth’s cells, enterocytes, goblet’s cells and neuroendocrine cells, respectively. Data represent average expression relative to NC non-differentiated ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, two-side unpaired t-test. ††p < 0.01, †††p < 0.001 Mann-Whitney test; 20 and 16 replicates for n = 4 non-celiac (NC) and n = 3 active celiac (CD) patients.
