| DMEM (Gibco/Life Technologies 11960-044) |
430 mL |
86% |
FCS (100%; heat-inactivated for 30 min at 55°C; Gibco/Life Technologies 10437-028)
|
50 mL |
10% |
Penicillin-streptomycin (100×; Gibco/Life Technologies 15140-122)
|
5 mL |
1× |
l-glutamine (200 mm; Gibco/Life Technologies 25030-081)
|
5 mL |
2 mm |
Sodium pyruvate (100 mm; Gibco/Life Technologies 11360-070)
|
5 mL |
1 mm |
| Insulin stock (0.5 mg/mL)
|
5 mL |
5 µg/mL |
| NAC stock (5 mg/mL)
|
0.5 mL |
5 µg/mL |
Hydrocortisone stock (10 mm in DMSO; Sigma-Aldrich H0888)
|
0.5 mL |
10 µm |
|
|
|
| Filter-sterilize through a 0.22-μm filter. The final volume is 500 mL. Store at 4°C in the dark. |