DMEM-Sato Base Growth Medium

1. Combine the following:

Reagent	Amount (for 20 mL)	Final
Dulbecco’s Modified Eagle’s Medium (Invitrogen 11960-044)	19.5 mL	1×
Sato supplement (100×)	200 µL	1×
Glutamine (200 mm; Invitrogen 25030-081)	200 µL	2 mm
Penicillin-streptomycin (Gibco/Life Technologies 15140-122)	200 µL	100 U/mL (penicillin)100 µg/mL (streptomycin)
Sodium pyruvate (100 mm; Invitrogen 11360-070)	200 µL	1 mm
Insulin stock (0.5 mg/mL)	200 µL	5 µg/mL
N-Acetyl-l-cysteine (Sigma-Aldrich A8199) stock (5 mg/mL, prepared in DMEM)	20 µL	5 µg/mL
Trace Elements B (1000×; Cellgro 99-175-CI)	20 µL	1×
d-Biotin (Sigma-Aldrich B4639) stock (50 µg/mL)	4 µL	10 ng/mL

2. For mouse OPCs only, include 400 µL of 50× B-27 (Invitrogen 17504-044).
- Commercially available B-27 contains small but biologically relevant amounts of T3. If you wish to study OPCs in the absence of T3-mediated signaling, you should use an alternative custom supplement called NS21, omitting the T3 (Chen et al. 2008), or use rat OPCs, which do not require B-27 for survival.
3. Filter through a 0.22-µm filter to sterilize. Store at 4°C for up to 1 wk.

REFERENCE