NPC medium was aspirated and NPCs were incubated with 1 ml/well of Accutase for 5–10 min at 37 °C. Once detached, 2 ml of DMEM was added per well, and NPCs transferred to a 15 ml tube containing DMEM and centrifuged (5 min × 1000g). Following aspiration and resuspension in NPC medium, cell concentration was determined with a hemocytometer. Typically, 2.5–5.0 × 105 NPCs were seeded into each well of a 24-well and evenly distributed. Cells were incubated overnight.
