Fig. 5: Intestinal sub-epithelial myofibroblasts (IMFs) act as a crucial extrinsic niche factor in small intestinal organoid architecture/organisation.

a Schematic shows the fbxw7^fl/fl before and after Cre recombination to generate fbxw7 gut-specific inactivation (fbxw7^ΔG) mice. Lower panels: ISH for fbxw7 and olfm4 mRNA on intestinal sections of 3-week fbxw7^fl/fl (left) and fbxw7^ΔG (right) mice. Scale bars, 50 μm. b Morphological representative images of a 7-day time course of small intestinal organoid growth from a single crypt isolated from fbxw7^fl/fl (left panels) and fbxw7^ΔG mice (right panels). Dashed lines indicate erupted epithelial cells from the fbxw7^ΔG crypts. Scale bars, 25 μm. c–f Graphs report the percentage of different morphologies found within a population of fbxw7^fl/fl, fbxw7^ΔG, Ep^ΔG:IMF^fl/fl (fbxw7^ΔG organoids seeded on a layer of wild-type intestinal myofibroblasts) and Ep^ΔG:IMF^ΔG (fbxw7^ΔG organoids seeded on a layer of fbxw7^ΔG-derived myofibroblasts) organoids cultured for 1 week. Organoids were classified as enterospheres (spherical structures), enteroids (lumens and budding development with multilobulated structures), microadenoma-like structures and spheres (organoids with 1–4 small buddings). Data are from four mice per genotype with the same sex and show mean% changes over the total number of organoids in co-cultures of crypt epithelial cells and myofibroblasts (Ep:IMF), compared with a single culture of crypt epithelial cells (Ep) ± standard error of the mean (SEM) for n = 4 parallel wells/condition. Error bars represent SEM; (*) value Ep^fl/fl vs. Ep^ΔG and (^o) value Ep^ΔG:IMF^fl/fl vs. Ep^ΔG:IMF^ΔG, ***P or ^oooP ≤ 0.001; **P or ^ooP ≤ 0.01; *P or ^oP ≤ 0.05, as determined by Student’s t test