Complete Growth Medium
ACL-4 medium (serum-free)
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 (final conc.)
- 50 nM Hydrocortisone (final conc.)
- 1 ng/ml Epidermal Growth Factor (do not filter)
- 0.01 mM ethanolamine (final conc.)
- 0.01 mM phosphorylethanolamine (final conc.)
- 100 pM triiodothyronine (final conc.)
- 0.5% (w/v) bovine serum albumin (final conc.)
- 10 mM HEPES
- 0.5 mM sodium pyruvate (final conc.)
- extra 2mM L-glutamine (for final conc. of 4.5mM)
Cultures can be maintained by the addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 x 106 viable cells/mL. Maintain cell density between 1 x 106 and 3 x 106 cells/mL.
Medium Renewal: Add fresh medium every 3 to 4 days (depending on cell density).
Culture medium, 85%; fetal bovine serum, 10%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.