| Dulbecco's modified Eagle's medium (DMEM) |
218 mL |
| Ham's F12 (Life Technologies 11765-054) |
72 mL |
| l-Glutamine (200 mm; Cellgro 25-005CI)
|
6 mL |
| Hydrocortisone (Sigma-Aldrich H0888) |
600 µL |
| ITES supplement (500×; Lonza 17839Z) |
600 µL |
| O-Phosphorylethanolamine (Sigma-Aldrich P0503)
|
600 µL |
| Adenine (Sigma-Aldrich A9795) |
600 µL |
| Progesterone (Sigma-Aldrich P8783) |
600 µL |
| Triiodothyronine (Sigma-Aldrich T5516) |
600 µL |
| Newborn calf serum (NBCS) (HyClone SH3011802) |
300 µL |
| Gentamicin sulfate (50 mg/mL; Cellgro 30-005-CR) |
300 µL |
| The final volume is sufficient for 12 inserts. |