Figure 4.

Apical-Out Enteroids Proliferate and Differentiate

(A) Basal-out enteroids and apical-out enteroids have proliferating cells when cultured in growth media (Growth), but not in differentiation media (Diff).

(B) Quantification of proliferation marker Ki67 shows that both basal-out and apical-out enteroid cells proliferate less in differentiation media (Diff) than in growth media (Growth). The percentage of proliferating cells in apical-out enteroids in growth media decreases over time. Data represented are mean ± SD; n = 40 enteroids; ^∗p < 0.05; ^∗∗∗∗p < 0.0001; NS, not significant using the Kruskal-Wallis test with Dunn’s multiple comparison test.

(C) Markers for different epithelial cell types (lysozyme for Paneth cells, MUC2 for goblet cells, CHGA for entero-endocrine cells, and villin for enterocytes) are expressed in apical-out enteroids.

Nuclei in blue and actin in white are shown. All scale bars are 10 μm. See also Figures S5 and S6.