Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended Medium Renewal: Every 2 to 3 days Remove spent medium, add fresh 0.25% trypsin, 0.03% EDTA solution, rinse and remove trypsin. Allow the flask to sit at room temperature, or incubate at 37C, until the cells detach (about 5 to 10 minutes). Add fresh medium. |