We describe the method for efficiently differentiating human embryonic stem cells to neuroepithelial cells in a chemically defined condition. The protocol was established based on the fundamental principle of in vivo neuroectodermal development. The temporal course, morphological transformation, and shift in gene expression of our neuroepithelial differentiation closely resemble those occur during in vivo development. In particular, the primitive neuroepithelial cells generated by this protocol can be further induced into neuronal and glial cells with forebrain, mid/hind brain, and spinal cord identities and targeted transmitter phenotypes.
