Complete Growth Medium
Dulbecco's modified Eagle's medium with 0.25 mg/ml xanthine, 0.015 mg/ml hypoxanthine, 0.01 mg/ml thymidine, 0.002 mg/ml aminopterin and 0.015 mg/ml mycophenolic acid, 90%; fetal bovine serum, 10%
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Remove medium, add fresh 0.25% trypsin, 0.02% EDTA solution for 1 to 2 minutes, rinse and remove trypsin.
Allow the cells to sit at 37C until they begin to detach (about 3 to 5 minutes), add fresh medium, aspirate and dispense into new flasks.
Subculture before the cells become confluent.
If allowed to become confluent, the cells will slough off and die.