Fig. 2: Exogenous Wnt3a micropatterns regulate crypt-like domain size and shape.

a Representative images of cells immunostained for Lysozyme (Lyz), GFP, F-Actin and nuclei (DNA) (upper panels), cell area segmentation and contour map (lower panels). Scale bars: 50 μm. b Cell area distributions in the crypt- and villus-like domains. Symbols correspond to experimental measurements, and solid lines are the Gaussian fits. Mean ± SEM. N = 4 experiments. c Representative images of Wnt3a micropatterns after cell seeding and growth (upper panels), cell area segmentation and cell area contour map (lower panels). Scale bars: 50 μm. d Bars correspond to experimental measurements of crypts’ area on Wnt3a patterns, and the dashed line corresponds to the distribution on control surfaces (Lognormal fit). Mean ± SEM. N = 3 experiments. e Bars correspond to experimental measurements of crypts’ circularity on Wnt3a patterns, and the dashed line corresponds to the distribution on control surfaces (Gaussian fit). Mean ± SEM. N = 3 experiments. f Representative images of EphrinB1 micropatterns after cell seeding and growth (left panels), cell area segmentation and area contour map (right panels). Scale bars: 50 μm. g Bars correspond to experimental measurements of crypts’ area on EphrinB1 patterns, and the dashed line corresponds to the distribution on control surfaces (Lognormal fit). Mean ± SEM. N = 3 experiments. h Bars correspond to experimental measurements of crypts’ circularity on EphrinB1 patterns, and the dashed line corresponds to the distribution on control surfaces (Gaussian fit). Mean ± SEM. N = 3 experiments. i Organoid-derived single cells were seeded on Wnt3a micropatterns of different sizes after 48 h of culture. Representative fluorescence images of F-Actin, DNA and Wnt3a (upper panels). Representative cell area contour maps (lower panels). The diameters of the Wnt3a micropatterns are 100 μm (middle panel) and 200 μm (right panel). Scale bars: 100 μm. j Crypt-like domain areas. Mean ± SEM. One-way ANOVA. N_control = 6, N₁₀₀ = 3 and N₂₀₀ = 3 experiments. k One-way ANOVA. N_control = 6, N₁₀₀ = 3 and N₂₀₀ = 3 experiments. Source data are provided as Source Data file.