Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: 2 to 3 times per week
Remove spent medium, add fresh 0.25% trypsin for 3 to 5 minutes, remove trypsin and let the culture sit at room temperature for 10 to 20 minutes. Add fresh medium, aspirate and dispense into new flasks. Subculture every 5 to 7 days.