StemFlex medium + 10 μM Y-27632 + 1:100 Pen/Strep
Media composition for differentiation
D0-1 - DMEM/F-12+Glutamax + 13 B-27 minus vitamin A + 13 N-2 + 100 nM LDN193189 + 10 μM SB431542
D2-3 - DMEM/F-12+Glutamax + 13 B-27 minus vitamin A + 13 N-2 + 100 nM LDN193189 + 10 μM SB431542+ 2 μM Purmorphamine + 1 μM SAG
D4-7 - DMEM/F-12+Glutamax + 13 B-27 minus vitamin A + 13 N-2 + 100 nM LDN193189 + 10 μM SB431542
+ 2 μM Purmorphamine + 1 μM SAG + 3 μM CHIR99021
D8-11 - DMEM/F-12+Glutamax + 13 B-27 minus vitamin A + 13 N-2 + 100 nM LDN193189 + 3 μM CHIR99021
D12-24 (Terminal Media) DMEM/F-12 + Glutamax + 13 B-27 minus vitamin A + 13 N-2 + 20 ng/mL BDNF + 20 ng/mL GDNF + 0.2 mM Ascorbic Acid + 10 μM DAPT + 0.1 μM dcAMP
D25+ (Long-Term Media) DMEM/F-12+Glutamax + 13 B-27 minus vitamin A + 13 N-2 + 10 ng/mL BDNF + 10 ng/mL GDNF + 0.2 μM Ascorbic Acid + 10 ng/mL CNTF.