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 1 to 2 minutes, remove trypsin and let the culture sit at room temperature for 5 to 10 minutes. Add fresh medium, aspirate and dispense into new flasks. Subculture every 6 to 8 days.