Complete Growth Medium
Minimum essential medium (Eagle) with 2 mM L-glutamine and Earle's BSS adjusted to contain 1.5 g/L sodium bicarbonate, 0.1 mM non-essential amino acids, and 1.0 mM sodium pyruvate, 90%, fetal bovine serum, 10%
Subculturing
Remove spent medium, add fresh 0.25% trypsin, 0.03% EDTA solution, rinse and remove trypsin. Let the culture sit at room temperature (or at 37°C) for 2 to 5 minutes. Add fresh medium, aspirate and dispense into new flasks. Subculture every 6 to 8 days.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: Every 3 to 4 days
