Fig. 5: Directed differentiation of intestinal organoids via niche signaling modulation.

a Schematic of small molecules screening strategy for induction of differentiated cell types. Differentiation protocols for Paneth cells (b), goblet cells (c), enteroendocrine cells (d), and enterocytes (e) (upper panels) and representative confocal images of organoids after differentiation showing Paneth cells (LYZ), goblet cells (MUC2), enteroendocrine cells (CHGA) and enterocytes (ALPI) (lower panels). Representative images from three independent experiments. f–i Quantification of the percentage of differentiated cells as shown in (b–e) respectively. One-way ANOVA with Tukey’s multiple comparisons test; data are presented as mean ± SD; representative experiment showing n = 3 samples from each condition. j Schematic of differentiation conditions for organoids cultured in TpC and TpCI (iBET-treated) conditions for experiments (k and l). k Representative confocal images of differentiated organoids, showing Paneth cell (LYZ), goblet cell (MUC2), and enteroendocrine cells (CHGA) in TpC and TpCI organoids. Representative images from three independent experiments. l Quantification of the percentage of differentiated cells as in (k). Two-tailed unpaired t-test; data are presented as mean ± SD; representative experiment showing n = 3 samples from each condition. m Schematic and representative confocal images illustrating M cell differentiation in organoids cultured under TpC condition. Representative images from three independent experiments. Scale bars, 50 μm. Source data for this Figure are provided as a Source Data file.