Disclosed herein are methods for differentiation of neuronal cells from stem cells, which produce a three-dimensional model of the neocortex structure. In some embodiments, the methods include culturing stem cells with at least one inhibitor of SMAD signaling and at least one inhibitor of fibroblast growth factor signaling for a period of time to result in formation of an embryoid body. The embryoid body is then cultured with at least one FGF (for example, basic FGF) for a period of time sufficient to produce a neuroepithelial (NE) rosette. The rosette is then isolated and cultured in suspension or adherent culture in the presence of at least one FGF for a period of time sufficient to produce cells with dorsal cortical identity and those cells are then cultured in the substantial absence of trophic factors for a period of time sufficient to form a neocortical organoid.
Three-dimensional model of human cortex