Complete Growth Medium
|
Dulbecco's modified Eagle's medium with 4 mM L-glutamine modified to contain 4.5 g/L glucose, 1.5 g/L sodium bicarbonate and 1 mM sodium pyruvate, 90%; fetal bovine serum, 10%
|
Subculturing
|
Remove medium, and rinse the monolayer with fresh 0.25% trypsin, 0.53 mM EDTA solution. Remove the trypsin, add fresh trypsin and let the culture sit at room temperature (or at 37°C) until the cells detach (about 10 minutes). Add fresh medium, aspirate and dispense into new flasks.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: Twice per week
|
Cryopreservation
|
Freeze Medium: Culture medium, 95%; DMSO, 5%
Storage Temperature: Liquid nitrogen vapor phase
|