Figure 1.

Comparison of 2D versus 3D In Vitro Cell Cultures

(A) Experimental design. TD, terminal differentiation day; EB, embryoid bodies; MON, monolayer; ORG, organoid.

(B) Representative images of immunocytochemical staining with the dorsal telencephalic marker PAX6, the neuroectodermal marker SOX1, the proliferative marker Ki67, and the neuron-specific class III β-tubulin at TD11.

(C) Proportion of SOX1⁺ and PAX6⁺ cells by stereological quantification over DAPI⁺ nuclei.

(D) Immunocytochemical staining of excitatory (TBR1⁺ and CTIP2⁺) and inhibitory (GABA⁺, GAD67⁺) cortical neurons at TD31.

(E) Proportion of excitatory and inhibitory neurons over DAPI⁺ nuclei assessed by stereological analysis. Results in (C and E) are the mean ± SEM of n = 3 biologically different iPSC lines per condition (ORG, MON) differentiated in parallel with two technical replicates per cell line.

^∗∗p < 0.01, ^∗p < 0.05 Student's t test, two tailed. See also Figure S1.