Figure 1.

Morphological Heterogeneity Is Robust in Normal Organoid Cultures

(A) A stitched micrograph of a Matrigel dome that embeds normal colonic organoids cultured for 6 days (left). A bull's eye (dotted circles) divided into three regions was used for the quantification. The core, intermediate, and edge regions are indicated with light, pale, and dark blue colors, respectively. High-magnification images focusing on the core (right top) and edge (right bottom) display significant differences in organoid size.

(B) The size distribution of normal human intestinal organoids across the entire matrix dome area corresponding to the stitched image in “A” (left) and the mean organoid size in each region (right). A total of four Matrigel domes that contain normal intestinal organoids were analyzed to estimate the cross-sectional area of individual organoids using ImageJ. The color in each column is matched to the corresponding location in (A).

(C) A stitched image of a Matrigel dome that contains CRC organoids cultured for 6 days under the same culture condition provided in (A) (left). High-power magnified images confirm that the CRC organoids in the core (right top) are morphologically similar to those in the edge (right bottom).

(D) The size distribution of CRC organoids across the dome area corresponding to the stitched image in (C) (left), and the mean organoid size in each region (right). Similar to the normal organoid, four Matrigel domes that contain CRC organoids were analyzed to obtain the cross-sectional area of individual CRC organoids.

A color bar at the bottom in both (A) and (C) shows the matched region in (B) and (D). Scale bars, 1 mm in the left images in both (A) and (C) and 200 μm in the right insets in both (A) and (C). Data are represented as mean ± SEM. One-way ANOVA with multiple comparison was performed to evaluate statistical significance of the differences. ∗p < 0.05; ns, not significant.