Complete Growth Medium
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
To make a 500 mL volume:
- mouse Interleukin-3 culture supplement (BD Biosciences, Catalog No. 354040) 25% (final conc.)
- fetal bovine serum 10% (final conc.)
- 325 mL DMEM (ATCC 30-2002)
- 125 mL Mouse Interleukin-3 (BD 354040)
- 50 mL FBS (ATCC 30-2020)
This medium is formulated for use with a 5% CO2 in air atmosphere. Standard DMEM formulations contain 3.7 g/L sodium bicarbonate. A 10% CO2 in air atmosphere is recommended.
Cultures can be maintained by addition or replacement of fresh medium. Start cultures at 2 X 105 cells/mL and maintain between 1 X 105 and 1 X 106 cells/mL.
Medium Renewal: Every 2 to 3 days
Freeze medium: Complete growth medium, 90%; DMSO, 10%
Storage temperature: liquid nitrogen vapor temperature