Complete Growth Medium
RPMI 1640 medium - OR - Minimum essential medium (Eagle) in Earle's BSS with non-essential amino acids, 90%; fetal bovine serum, 10%
Remove medium, add fresh 0.25% trypsin or trypsin, versene for 2 to 3 minutes, remove trypsin and allow the culture to sit at room temperature for 5 to 10 minutes.
Add fresh medium, aspirate and dispense into new culture vessels.
Subculture every 7 to 10 days.
Medium Renewal: 2 to 3 times per week
Subcultivation Ratio: A subcultivation ratio of 1:3 (or 2 X 10 exp6 cells per 75 sq cm flask) is recommended