Table S3.

Media and additives used in this study

Medium/additive	Composition (final concentration)	Notes
Base media
Serum	DMEM (Gibco, 11995–065); 15% FBS (Gibco, 10270–106); 1× pen/strep solution (Gibco, 15140–122); 1× nonessential amino acids (Gibco, 11140–035); 50 nM β-mercaptoethanol (Gibco, 31350–010)
N2B27	1:1 DMEM/F12 (Gibco 21331):Neurobasal (Gibco 21103); 1× N2 (Gibco, 17502–048); 1× B27* (Gibco 17504–044 or 12587–010); 0.4 mM l-glutamine (Gibco 25030–024); 1× pen/strep solution (Gibco, 15140–122); 50 nM β-mercaptoethanol (Gibco, 31350–010)	*B27 with retinyl acetate (VitA+) is 17504–044; B27 without retinyl acetate (VitA−) is 12587–010
Additives
2i	3 µM, CHIR99021 (Stemgent, 04–0004); 1 µM PD0325901 (Stemgent, 04–0006)
LIF	10 μg/mL LIF (Wellcome Trust Centre for Stem Cell Research)
FGF2	12.5 ng/mL FGF2 (Wellcome Trust Centre for Stem Cell Research)
Activin A	20 ng/mL Activin A (Wellcome Trust Centre for Stem Cell Research)
Blasticidin	20 μg/mL Blasticidine S hydrochloride (Sigma-Aldrich, 15205)
Retinol	Variable concentrations (Sigma-Aldrich, R7632)
Retinoic acid	1 mg/mL (Sigma-Aldrich, R2625)
Ascorbate	50 μg/mL (Sigma-Aldrich, A7631)