Cell -Freeze Down

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Freeze Media

9ml media the cells grow in

1ml DMSO

  1. Trypsinize as normally.
  2. Spin cells and resuspend in appropriate freeze media for ~1-2million cells/vial.
  3. Place 1ml of Freeze Media.
  4. Place cryovial in Nalgene cell freezer and put at -80°C until next day. (If you do not use a Nalgene cell freezer, this protocol will NOT work)
  5. Transfer the cryovials into liquid nitrogen until use.

To thaw- place frozen cryovial in to 37oC water bath for 2-3 minutes. Using sterile forceps remove tissue fragments and rinse in sterile PBS. Tissues are now ready for use or implantation.