Basal medium		Comments
Store at 4 °C for 4 weeks
Cell culture medium	500 mL	See table of materials
L-Glutamine 100x	5 mL
Buffering agent 1 M	5 mL	See table of materials
Penicillin Streptomycin 100x	5 mL
WENR + Nic (Wnt + EGF + Noggin + R-spondin + Nicotinamide)
Store at 4 °C for 2 weeks
Basal medium	up to 50 mL
Neuronal cell serum-free supplement (50x)	1 mL	See table of materials
Neuronal cell serum-free supplement (100x)	500 μL	See table of materials
n-Acetylcysteine (500 mM)	125 μL
mouse EGF (100 μg/mL)	25 μL
mouse Noggin (100 μg/mL)	50 μL
R-Spondin conditioned medium	5 mL
Wnt3a conditioned medium	25 mL
Nicotinamide (1 M)	250 μL
EN + CHIR + Y-27632 (EGF + Noggin + CHIR + Y-27632)
Store at 4 °C for 2 weeks
Basal medium w/o Penicillin Streptomycin	up to 20 mL
Neuronal cell serum-free supplement (50x)	400 μL	See table of materials
Neuronal cell serum-free supplement (100x)	200 μL	See table of materials
n-Acetylcysteine (500 mM)	50 μL
mouse EGF (100 μg/mL)	10 μL
mouse Noggin (100 μg/mL)	20 μL
Y-27632 (10 μM)	20 μL
CHIR99021 (8 μM)	10 μL
EN (EGF + Noggin)
Store at 4 °C for 4 weeks
Basal medium	up to 50 mL
Neuronal cell serum-free supplement (50x)	1 mL	See Table of materials
Neuronal cell serum-free supplement (100x)	500 μL	See Table of materials
n-Acetylcysteine (500 mM)	125 μL
mouse EGF (100 μg/mL)	25 μL
mouse Noggin (100 μg/mL)	50 μL