Composition	Volume (520 mL total)	Final concentration	Comments
Neurobasal medium	500 mL	–	M2 medium
GlutaMAX	5 mL of 100×	1×
NEAA	5 mL of 100×	1% (vol/vol)
Gem21	1 bottle (10 mL)	2% (vol/vol)
BDNF	–	10 ng/mL (1: 2,000 of 20 μg/mL stock)	Add just before use
GDNF	–	10 ng/mL (1: 2,000 of 20 μg/mL stock)	Add just before use
NT-3	–	10 ng/mL (1: 2,000 of 20 μg/mL stock)	Add just before use
Ascorbic acid	–	200 μM (1: 1,000 of 200 mM stock)	Add just before use
Dibutyryl-cAMP	–	1 mM (1: 200 of 200 mM stock	Add just before use