Structural Neural Network Development of the 3D In Vitro Biomimetic Cortical Culture
(A) Representative 3D projections of confocal z stack images of RFP-expressing neurons at 1, 2, and 3 weeks. Scale bar, 20 μm.
(B) Representative 3D rendering of side (x-z) view of RFP-expressing neurons at 3 weeks.
(C) Neurite density in the 3D cortical culture (mean ± SEM). n = 6–8 samples from three independent experiments.
(D) mRNA expression over time (mean ± SEM) of proteins associated with neural progenitors (Pax6), immature neurons (Dcx, doublecortin), presynaptic vesicles (Syn1, synapsin 1), postsynaptic scaffolding protein (Shank3), glutamatergic transmission (Slc17a7, vesicular glutamate transporter 1; Gria1, AMPA receptor subunit 1;Grin1, NMDA receptor subunit 1), and GABAergic transmission (GAD2, glutamate decarboxylase 2; Gabra1, GABAA receptor subunit ɑ1). Expressions were normalized to housekeeping gene 18s. n = 7–11 samples from three to four independent experiments. For (C) and (D), ANOVA and post-hoc Tukey tests were used for statistical significance among groups. ∗p < 0.05; ∗∗p < 0.001. See also Video S1. See also Table S2.