Protocol for Reviving CHO-R5 Cell Line
- Pipet 25 ml of growth media into a T75 flask. Add G418 to a final concentration of 500 µg/ml.
- Incubate for 30 minutes at 37°C, 5% CO2 to equilibrate media.
- Thaw the tube containing the frozen cells in a 37°C water bath for 2 minutes.
- Immediately transfer the thawed cell stock to the flask containing the equilibrated growth media.
- Incubate cells overnight at 37°C, 5% CO2 and replace media the next day.
- Continue to incubate the revived cells for 48 hours (they will reach confluence at about 2 days).