Fig. 5: Mechanically enforced cell multilayering and crowding are essential for enhancing the initiation of gut tissue budding via a buckling-like mechanism.

a (upper) Representative X-Z section showing the architecture of the gut tissue cultured in μGSG (micropatterned gut spheroid generator), with F-actin stained in green and nuclei in blue. The buckled configuration of F-actin cortex was outlined with the white dashed line. Similar results were seen in n = 3 independent experiments. (lower) Schematic of the tissue architecture shown in the upper panel, which was used to conduct finite element analysis of tissue buckling under crowding-induced compressive strain (see Methods for detailed descriptions). b Simulation results showing the change in normalized buckling amplitude as a function of the tissue-to-cortex thickness ratio (h_t / h_c). Results obtained under 1.2%, 2.5%, and 5% lateral compressive strain was shown, respectively. c Contours of tissue deformation under different levels of compressive strain for monolayer tissue (h_t = 15 μm) and multilayer tissue (h_t = 30 μm), respectively. Displacement field in the y direction (U₂) was color-coded as shown by the color bars. Source data are provided as a Source Data file.