days onto polyornithine/laminin (Sigma)-coated dishes in DMEM/F12 (Invitrogen) plus N2. Rosettes were manually collected and dissociated with Accutase (Chemicon) after 1 week and plated onto coated dishes with NPC media (DMEM/F12, N2, B27, and FGF2). To obtain mature neurons, NPCs were plated on a monolayer of hippocampal astrocytes in the presence of DMEM/F12, N2, B27, ascorbic acid (200 nM), cyclic AMP (cAMP; 500 μg/ml), laminin (1 μg/ml), BDNF (20 ng/ml), Wnt3a (20 ng/ml), and 1% fetal bovine serum for 3 weeks. Wnt3a was removed after 3 weeks.
