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. |
