iPSC Neural Progenitor Cell Differentiation

iPSCs (line 9319 from Brennand et al, 2011 (link)) were maintained under standard conditions and dissociated from plates using collagenase type IV (Invitrogen, Carlsbad, CA, USA) to form floating embryoid bodies, which were cultured in DMEM/F12 + glutamax (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 1% N2 (Thermo Fisher Scientific, Waltham, MA, USA), 2% B27 without vitamin A (Thermo Fisher Scientific, Waltham, MA, USA), SB 431542 (Tocris, Bristol, UK) and LDN193189 (Stemgent, Cambridge, MA, USA)). After 7 days, the embryoid bodies were re-plated onto polyornithine (Sigma- Aldrich,St. Louis, MP, USA) and laminin (Thermo Fisher Scientific, Waltham, MA, USA) coated plates and cultured in the same media for an addition 7 days until neural rosettes formed. Neural rosettes were dissociated using neural rosette selection reagent (Stemcell, Vancouver, Canada) and replated onto Corning™ matrigel (Fisher Scientific, Hamptom, NH, USA))-coated plates to form populations of neural progenitor cells (NPCs).