DEVELOPMENT OF NOVEL CRYOPRESERVATION TECHNIQUE FOR HUMAN RED BLOOD CELLS

WITH CURRENT FDA-APPROVED PROTOCOLS FOR THE CRYOPRESERVATION OF HUMAN RED BLOOD CELLS, THE CELLS ARE TREATED WITH MULTIMOLAR CONCENTRATIONS OF GLYCEROL, A PERMEATING CRYOPROTECTANT. AFTER THAWING, EXTENSIVE PROCESSING OF RED CELLS IS REQUIRED TO REMOVE THE GLYCEROL, PRIOR TO TRANSFUSION, TO PREVENT HEMOLYSIS. FURTHERMORE, EVEN IN THE PRESENCE OF CRYOPROTECTANTS, THE FORMATION OF ICE DURING COOLING AND ICE RECRYSTALLIZATION DURING WARMING ARE MAJOR DAMAGING EVENTS DURING FREEZE-THAWING OF RED BLOOD CELLS. TO CIRCUMVENT THESE OBSTACLES TO RED CELL PRESERVATION, WE WILL USE A NOVEL PROTEIN (ICE CRYSTAL CONTROL PROTEIN, ICCP) THAT DEPRESSES THE FREEZING TEMPERATURE OF WATER, MODIFIES ICE CRYSTAL SHAPE AND INHIBITS RECRYSTALLIZATION. ICCP, IN MICROMOLAR CONCENTRATIONS, WILL BE COMBINED WITH CONVENTIONAL CRYOPROTECTANTS (HYDROXYETHYL STARCH, GLYCEROL) AT REDUCED CONCENTRATIONS, THEREBY ELIMINATING THE NEED TO REMOVE THE CRYOPROTECTANT AFTER THAWING AND ALLOWING IMMEDIATE TRANSFUSTION. THESE PROTEINS ARE LIKELY TO BE NON-TOXIC TO HUMANS; THEY OCCUR NATURALLY IN POLAR FISHES AND INSECTS. CRYO-LIFE HAS EXCLUSIVE ACCESS TO ICCP, WHICH CAN BE PRODUCED IN INDUSTRIAL QUANTITIES BY GENETIC ENGINEERING, FOR THE CRYOPRESERVATION OF MAMMALIAN CELLS AND TISSUES.