File:Human telencephalic organoids are larger with extended apical lumens.jpg

English: "(A) Schematic of the timeline for generating brain organoids from human, gorilla, and chimpanzee stem cells. Colors represent changes in media and stages of the protocol. +MG represents Matrigel embedding. Note chimpanzee organoids have an EB stage that is 2 days shorter than human and gorilla.

(B) Bright field images of human (H9, top panels), gorilla (G1, middle panels), and chimpanzee (Chmp, bottom panels) organoids at days 3, 5, and 10. Black arrows indicate neuroepithelial buds, which appear more elongated in the human beginning at day 5. Scale bar, 200 μm. (C) Quantification of bright field images at day 10 show human (H9) neural tissue is enlarged relative to gorilla (G1) and chimpanzee (Chmp). Left: area occupied by individual organoids. Right: visible perimeter of individual neuroepithelial buds. Mean organoid area: H9 = 168,327 μm2; G1 = 84,876 μm2; Chmp = 81,086 μm2. Mean neuroepithelial bud perimeter: H9 = 322 μm; G1 = 237 μm; Chmp = 204 μm. ∗p < 0.05, ∗∗∗∗p < 0.0001, Kruskal-Wallis and post hoc Dunn’s multiple comparisons test, n (H9) = 22 organoids and 144 neuroepithelial buds from 6 independent batches, n (G1) = 23 organoids and 148 neuroepithelial buds from 5 independent batches, n (Chmp) = 13 organoids and 107 neuroepithelial buds from 3 independent batches, error bars are SD. (D and E) Representative immunofluorescence images of the center of whole mount human (H9), gorilla (G1), and chimpanzee (Chmp) organoids with staining for apical marker ZO1 and neural progenitor marker SOX2 at day 3 (D) and day 5 (E). Note the appearance of less rounded ZO1 positive apical lumens (arrowheads) in human organoids relative to nonhuman ape organoids at day 5. DAPI is in blue. Red background signal outside the organoid comes from nonspecific uneven staining of surrounding Matrigel. Scale bar, 100 μm. (F) 3D MATLAB reconstructions of apical lumens of day 5 organoids showing a representative example used for quantifications in (G). Values on the axes are in μm. Luminal surface area of reconstructed examples: human (H9) = 63,394 μm2; gorilla (G1) = 15,146 μm2; chimpanzee (Chmp) = 19,730 μm2.

(G) Quantification of the surface area of the largest apical lumen per day 5 organoid reveals significantly expanded luminal surface areas in human versus nonhuman apes. Mean luminal surface area: human (H9) = 51,243 μm2; gorilla (G1) = 15,437 μm2; chimpanzee (Chmp) = 19,632 μm2. ∗p < 0.05, ∗∗p = 0.0012, ∗∗∗∗p < 0.0001, one-way ANOVA and post hoc Tukey’s multiple comparisons test, n (H9 and G1) = 11 organoids from 5 independent batches, n (Chmp) = 8 organoids from 2 independent batches, error bars are SD."