Complete Growth Medium
HITES medium supplemented with 5% fetal bovine serum
The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No.30-2006. To make the complete growth medium,add the following components to the base medium
- 0.005 mg/ml Insulin
- 0.01 mg/ml Transferrin
- 30nM Sodium selenite (final conc.)
- 10 nM Hydrocortisone (final conc.)
- 10 nM beta-estradiol (final conc.)
- extra 2mM L-glutamine (for final conc. of 4.5 mM)
- 5% fetal bovine serum (final conc.)
Remove medium, and rinse with 0.25% trypsin, 0.03% EDTA solution. Remove the solution and add an additional 1 to 2 mL of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37°C) until the cells detach. Add fresh culture medium, aspirate and dispense into new culture flasks.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Freeze medium: RPMI 1640, 85%; fetal bovine serum, 10%; DMSO, 5%.