Primary Tissue Isolation-Freeze Down

Freeze Media

9ml DMEM + 10% CS + P/S/F (antibiotics/anti fungals)

1ml DMSO

  1. Remove tissues and place in sterile Petri dish.
  2. Chop/mince tissue with STERILE razor blade into 1mm cubes.
  3. Place 4-5 tissue fragments into a cryovial containing 1ml of Freeze Media.
  4. 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)
  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.