Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended Medium Renewal: Every 3 to 4 days Remove spent medium, add fresh 0.25% trypsin, 0.03% EDTA solution, rinse and remove trypsin. Add fresh trypsin solution (1 to 2 ml) and let the culture sit at room temperature (or at 37C) until the cells detach. Add fresh medium, aspirate and dispense into new flasks. Subculture every 6 to 8 days. |