9ml media the cells grow in
- Trypsinize as normally.
- Spin cells and resuspend in appropriate freeze media for ~1-2million cells/vial.
- Place 1ml of Freeze Media.
- 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)
- 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.