In vitro neural differentiation culture of embryonic stem cells (ESCs) provides a promising tool for preparing neural cells for replacement therapies and a versatile system for understanding mechanisms of neurogenesis. Consistent with the neural-default model, neural differentiation spontaneously occurs in ESCs cultured in medium containing minimal extrinsic signals. Both adherent monolayer culture and floating aggregation culture can be used for ESC conversion into neural progenitors. The floating aggregation culture has an advantage for recapitulating the formation of three-dimensional (3D) neural tissue structure such as layer formation. In this article, we review recent progress in neural differentiation culture of ESCs using 3D culture, focusing on self-organization phenomena of stratified cortex and retinal tissues. These self-organizing processes are driven by both cell intrinsic programs and local cell-cell interactions. A simple in vitro system using ESCs is useful for elucidating mechanistic dynamics in the complex orchestration of neural development.
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