Neuronal Differentiation of NPCs

AD and HN1 NPCs were expanded with STEMdiff™ Neural Progenitor Medium (NPM) (cat no. 0583 STEMCELL Technologies) on 6-well plates coated with PLO and laminin. Full volume media changes were performed every second day. Cells were switched to STEMdiff™ Neural Induction Medium (NIM) (STEMCELL Technologies cat no. 05385) and were supplemented with 100 ng/mL purmorphamine (puro) (STEMCELL Technologies cat no. 72204) on days 1–6, 100 ng/mL puro and 100 ng/mL fibroblast growth factor 8 (FGF-8) (STEMCELL Technologies cat no. 78128) on days 6–12, and 100 ng/mL puro, 100 ng/mL FGF-8, and 10 ng/mL bone morphogenic protein 9 (BMP9) (Peprotech cat no. 120-07) on day 12 with half volume media changes every second day to prepare for bioprinting (Muñoz et al. 2020 (link)).