| DMEM-High Glucose medium (Gibco 11960-044) |
39 mL |
| Insulin stock (0.5 mg/mL)
|
400 µL |
| Sodium pyruvate (100 mm) (Gibco 11360-070)
|
400 µL |
| l-Glutamine (200 mm) (Gibco 25030-081)
|
400 µL |
| Hormone mix (200×)
|
200 µL |
| T3 stock (4 µg/mL) |
200 µL |
| Hydrocortisone (50 µm) (Sigma-Aldrich H6909)
|
40 µL |
| Trace Elements B (1000×) (Cellgro) |
40 µL |
| Biotin stock (5000×)
|
8 µL |
| Vitamin B12 (Sigma-Aldrich V6629; 1.36 mg/mL in D-PBS) |
8 µL |
| B-27 (Invitrogen) or N21-MAX (R&D Systems) |
15 µL |
| Ceruloplasmin (1 mg/mL) (Calbiochem 239799) |
4 µL |
| Filter the solution through a 0.22-µm filter to sterilize and store at 4°C for up to 1 wk. |