Three-Dimensional Gastrointestinal Organoid Culture in Combination with Nerves or Fibroblasts: A Method to Characterize the Gastrointestinal Stem Cell Niche

Figure 3

Fibroblasts improve organoid growth and organoid culture survival. (a) Macroscopic image of the crypt monoculture and coculture in an organotypic cell culture system at day 7. (b) Characterization of the crypt monoculture (left panel) and coculture (right panel) in an organotypic cell culture system at day 7 by immunohistochemistry staining: H&E (scale bar 100 μm), PAS (scale bar 100 μm), Ki-67 (scale bar 50 μm), and α-SMA staining (scale bar 100 μm), with adjacent magnification. (c) Organoid diameter measured on H&E slides from crypt monoculture and coculture in organotypic cell culture system after 2 and 7 days of culture. Each dot represents a single organoid. (one-way ANOVA with Bonferroni comparison). (d) Lower, example of a nonviable crypt in organotypic cell culture system, H&E staining. Scale bar, 25 μm. Upper, survival of crypts in an organotypic cell culture system, for the monoculture 169 crypts were quantified; for the coculture 213 crypts were quantified. , two-tailed -test.