DRG Base Medium

Recipe

Reagent	Volume	Final concentration
DMEM (Gibco 11960-044)	37 mL	50%
Neurobasal (Gibco 21103-049)	37 mL	50%
Insulin (0.5 mg/mL)	800 µL	5 µg/mL
Sodium pyruvate (Gibco 11360-070)	800 µL	1 mm
Penicillin/streptomycin (100×; Gibco 15140-122)	800 µL	1×
l-Glutamine (Gibco 25030-081)	800 µL	1 mm
SATO supplement, NB-based (100×)	800 µL	1×
T3 stock (4 µg/mL)	800 µL	40 ng/mL
NS21^a (50×)	1.6 mL	1×
NAC stock (5 mg/mL)	80 µL	5 µg/mL
Final volume	80 mL

Filter-sterilize the medium through a 0.22-μm filter and store at 4°C for up to 1 wk. Add growth factors (NGF, BDNF, NT-3, and forskolin) immediately before use.
^aAlternatively, use N21-MAX Media Supplement (R&D Systems AR008) or B-27 (Invitrogen 17504-044). If using NS21 or B27 that contains T3, it is unnecessary to add T3 to DRG Base Medium.