Fig. 1

Isolation of intestinal epithelial cells from small intestinal organoids A Macroscopic view of intestinal organoids. Bar: 10 mm. Inset: High-power view. Bar: 5 mm. B Phase-contrast micrograph of small intestinal organoids with submucosa. Scale bar: 500 µm. C Phase-contrast photomicrographs of appropriate (left) and inappropriate (right) 'intestinal organoids'. Organ-derived small intestinal epithelium and mucosal stroma can be obtained from the appropriate organoids that have thick cell layers and low light transmission. Bar: 500 μm. D Histology of small intestinal organoids with submucosa. HE stains. Scale bars: 200 (left), 50 (right) µm. E Schematic of small intestinal organoids with submucosa. F Phase-contrast micrographs of an organoid cut open and unfolded with scissors. Scale bars: 500 μm. G Three-dimensional small intestinal organoids were bisected to expose the submucosa and attached to the dish. Cells were spread on the dish. H Macroscopic view of the dissected organoids that are attached to dishes. I Phase-contrast micrograph of a bisected organoid (black dotted lines) on 3, 13, 21, and 33 days after adhesion. Scale bars: 500 µm. J Phase-contrast photomicrographs of proliferating intestinal epithelial cells on the mucosal stroma. Scale bar: 100 µm. K Immunocytochemistry of adherent organoids with an antibody to CDX2, an intestinal epithelial marker. Nuclei were stained with 4',6-diamidino-2-phenylindole dihydrochloride solution (DAPI). Scale bars: 100 µm