Media Components
| Additions | Stock Concentration | Final Concentration | Amount added to 500 ml medium | |
| Growth Factors | Insulin (I) | 1 mg/ml | 5 μg/ml | 2.5 mL |
| Hydrocortisone (H) | 1 mg/ml | 1 μg/ml | 500 μL | |
| EGF (E) | 10 μg/ml | 10 ng/ml | 500 μL | |
| Cholera Toxin | 100 μg/ml | 100 ng/ml | 500 μL | |
| Progesterone | 20 μg/ml | 31.4 ng/ml (10-7 M) | 786 μL | |
| Prolactin | 1 mg/ml | 5 μg/ml | 2.5 mL | |
| Estradiol | 20 μg/ml | 2.7 ng/ml (10-8 M) | 68 μL | |
| Lysophosphatidic Acid | 10 mM | 10 μM | 500 μL | |
| Anti-fungal | Fungizone (Amphotericin B) | 250 μg/ml | 2.5 μg/ml | 5 mL |
| Bacteria | Gentamicin | 50 mg/ml | 25 μg/ml | 250 μL |
| Anti-mycoplasma | Plasmocin (maintenance) | 25 mg/ml | 2.5 μg/ml | 50 μL |
| Serum Replacement Factors | Ethanolamine | 16.6 M | 5 mM | 155 μL |
| HEPES | 1 M | 10 mM | 5 mL | |
| Transferrin | 2.5 mg/ml | 5 μg/ml | 1 mL | |
| Triiodo Thyronine (T3) | 20 μg/ml | 10 nM (6.6 ng/ml) | 167 μL | |
| Sodium Selenite (Se) | 20 μg/ml | 50 nM (8.7 ng/ml) | 217 μL | |
| Bovine Serum Albumin | 20 g/L | 1 g/L | 25 ml | |
| Serum | Fetal Bovine Serum | 5% | 25 mL |
For media containing serum: Begin with Ham’s F-12 and add 5% FBS plus the anti-fungal, anti-bacterial and anti-mycoplasma agents (if needed). Add appropriate growth factors as needed for each individual cell line.
For serum-free media: Begin with Ham’s F-12 and add all of the serum-replacement factors, the anti-fungal, anti-bacterial and anti-mycoplasma agents (if needed), plus appropriate growth factors as needed for each cell line.
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