Methods and compositions are provided for the cryopreservation of human organs and tissues. In certain embodiments, Step 1 comprises perfusion with a vitrifiable cryoprotectant solution at a temperature above −10° C. for a time insufficient for the approximate osmotic equilibration of the organ with the solution, followed by cooling the organ to below −10° C. by perfusion with said solution at a reduced temperature. In certain embodiments, Step 2 comprises increasing the concentration of cryoprotectant further at a temperature from −10 to −40° C. In certain embodiments, Step 3 comprises cooling and vitrifying the organ, rewarming it, and perfusing the organ with a vitrifiable concentration of cryoprotectant whose temperature is either raised gradually or is held at ≧−15° C. Compositions are provided that allow safe organ perfusion with vitrifiable media at >−10° C. and almost complete avoidance of chilling injury at −20 to −25° C. and that allow slow warming after vitrification without freezing.
