Protocol: Remove medium, and rinse with 0.25% trypsin, 0.53 mM EDTA solution. Remove the solution and add an additional 1 to 2 ml of trypsin-EDTA solution. Allow the flask to sit at room temperature (or at 37C) until the cells detach.
Add fresh culture medium, aspirate and dispense into new culture flasks.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended
Medium Renewal: At the time of subcultivation (every 5 to 7 days)
Freeze medium: Complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase