Restoration from DMSO cryopreservation¶
Contributed by Martin Fitzpatrick, University of Birmingham, United Kingdom
Restoration of culture cells from storage at -80°C with 10% DMSO cryopreservation.
- For each preserved sample add 6ml media (RPMI+10% FCS) to a 15ml tube.
- Defrost frozen samples in water bath at 37’C. Rinse with EtOH to clean before transferring to tissue culture hood.
- Pasteur all cells from sample into 6ml media, transfer back and forth to rinse.
- Centrifuge 15ml tube at 1200rpm 6 minutes to pellet.
- Pour off supernatant and resuspend in 2ml media. Transfer into 25cm^2 flask.
- Incubate overnight.
- If cells are respirating effectively media will turn yellow. Add media up to 5ml to culture.