Cells were dissociated into a single cell suspension with TriplE (Invitrogen, 12604-013) or Accutase and 3×104 NPCs were plated on glass coverslips coated with 10 µg/mL laminin (Sigma, L2020) and 15 µg/mL poly-ornithine (Sigma, P4957) in Neurobasal media (Invitrogen, 21103-049) containing 200 µM ascorbic acid, 0.5 mM dibutyryl cAMP (Sigma, D0260), and 1 mg/mL laminin, 20 ng/mL BDNF (Preprotec, 450-02), 20ng/mL GDNF (Preprotec, 450-10). Cells were fed every two to three days until ready for immunostaining and/or electrophysiology.
