Generation of iPSC-derived Neural Cells

Fibroblasts from neurotypical individuals were reprogrammed to induced pluripotent stem cells using lentiviral (9429) or non-integrating Sendai (Invitrogen) vectors^{12 (link)}. NPCs and neurons were generated following methods outlined by Brennand and Livesey^{91 (link),97 (link)}. Briefly, hiPSC-derived NPCs were plated on Matrigel (Fisher) or Poly-Ornithine (Sigma)/ Laminin (Invitrogen) and passaged 1:6 every 4 days. NPCs (< passage 14) were cultured in NPC medium (DMEM/F12 + Glutamax (Invitrogen), 1 × N2 (Invitrogen), 1X B27-Vitamin A (Invitrogen), 1ug/ml Laminin (Invitrogen), 20 ng/ml FGF-2 (Peprotech) and dissociated in Accutase (Fisher) for 5 min at 37 °C. hiPSC-derived neurons were cultured on Matrigel in Neuron Medium (DMEM/F12 + Glutamax, 1 × N2, 1X B27 with Vitamin A (Invitrogen), 20 ng/ml BDNF (Shenandoah Biotechnology), 20 ng/ml GDNF (Shenandoah Biotechnology), 1 mM dibutryl-cyclic AMP (Sigma), and 200 nM ascorbic acid (Stem Cell Technologies).