Complete Growth Medium
ACL-4 medium supplemented with 10% FBS
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:
- 0.02 mg/ml insulin
- 0.01 mg/ml transferrin
- 25 nM sodium selenite
- 50 nM Hydrocortisone
- 1 ng/ml Epidermal Growth Factor (do not filter)
- 0.01 mM ethanolamine
- 0.01 mM phosphorylethanolamine
- 100 pM triiodothyronine
- 0.5% (w/v) bovine serum albumin
- 10 mM HEPES
- 0.5 mM sodium pyruvate
- 2mM L-glutamine(for final conc. of 4.5 mM)
- 10% FBS
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
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 37C) until the cells detach.
Centrifuge, resuspend cells in fresh medium and dispense into new flasks.
RPMI 1640 medium 85%; fetal bovine serum, 10%; DMSO, 5%