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.