On D24 of neural induction, cultured cells were dissociated with accutase to allow clumps of 10–50 cells to remain and plated at a density of 8 × 105 cells/cm2 onto poly-D-lysine (Millipore) and laminin (Invitrogen)-coated 35mm and 24-well plates in N2/B27 + 2 μM thiasovivin.
