Complete Growth Medium
These cells are grown in Dulbeccos Modified Eagles Medium/Hams Nutrient Mixture F-12, 1:1 with 2.5 mM L-glutamine, 1.2 g/L sodium bicarbonate, 15mM HEPES and 0.5mM sodium pyruvate.
- 10% fetal bovine serum
- 10 µg/ml insulin
- 5.5 µg/ml transferrin
- 5 ng/ml selenium
- 40 ng/ml dexamethasone
This medium is formulated for use with a 5% CO2 in air atmosphere.
ATCC modified and tested medium formulation (without the additional supplements or serum described above) is available as ATCC 30-2006.
ATCC tested fetal bovine serum is available as ATCC 30-2020 (500ml) and ATCC 30-2021 (100ml).
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended
Medium Renewal: Every 2 to 3 days
Remove spent medium, add fresh 0.25% trypsin, 0.03% EDTA solution, rinse and remove trypsin.
Allow the flask to sit at room temperature, or incubate at 37C, until the cells detach (about 5 to 10 minutes).
Add fresh medium, aspirate and dispense into new flasks.