9ml DMEM + 10% CS + P/S/F (antibiotics/anti fungals)
- Remove tissues and place in sterile Petri dish.
- Chop/mince tissue with STERILE razor blade into 1mm cubes.
- Place 4-5 tissue fragments into a cryovial containing 1ml of Freeze Media.
- Place cryovial in Nalgene cell freezer and put at –80oC 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.