Radial glia and astrocyte differentiation from human pluripotent stem cells

A method of producing, in culture, radial glia-like cells, the method comprising:

• (a) plating vertebrate pluripotent stem cells on a substrate-coated surface of a culture vessel at a density of 1,000-100,000 cells/cm2;
• (b) incubating the plated vertebrate pluripotent stem cells in a first culture medium
• (c) replacing the first culture medium with a second culture medium comprising:
• (i) an effective amount or concentration of one or more inhibitors of BMP pathway, wherein the one or more inhibitors of the BMP pathway comprise one or more LDN-193189, LDN-214117, LDN-212854, DMH2, ML 347, UK 3833687, K 02288, Dorsomorphin, Noggin, Chordin, Follistatin, or Gremlin,
• (ii) an effective amount or concentration of one or more activators of Notch pathway wherein the effective amount or concentration of the one or more activators of Notch pathway in the second culture medium comprise one or more of Jagged 1 protein, Jagged 2 protein, and Delta-Like protein 1 (DLL1), Delta-Like protein 2 (DLL2), or Delta-Like protein 3 (DLL3), and
• (iii) one or more cytokines of interleukin-6 family;
• and
• (d) culturing the plated vertebrate pluripotent stem cells in the second culture medium;
• wherein the resulting radial glia-like cells detectably express one or more of Brain Lipid Binding Protein (BLBP), abnormal spindle-like microcephaly-associated protein (ASPM), baculoviral inhibitor of apoptosis repeat-containing 5 (BIRC5 or Survivin), FAT Atypical Cadherin 1 (FAT1), Hes family bHLH transcription factor 5 (HES5), and SRY-Box Transcription Factor 21 (SOX21).