Protocol for Reviving CHO-R5 Cell Line

Protocol

1. Pipet 25 ml of growth media into a T75 flask. Add G418 to a final concentration of 500 µg/ml.
2. Incubate for 30 minutes at 37°C, 5% CO₂ to equilibrate media.
3. Thaw the tube containing the frozen cells in a 37°C water bath for 2 minutes.
4. Immediately transfer the thawed cell stock to the flask containing the equilibrated growth media.
5. Incubate cells overnight at 37°C, 5% CO₂ and replace media the next day.
6. Continue to incubate the revived cells for 48 hours (they will reach confluence at about 2 days).