After storage in the liquid state at 4 C for up to three weeks, washing with sodium chloride solutions, and storage in a sodium chloride-glucose-phosphate solution for 24 hours at 4 C, dog red blood cells had excellent post-transfusion survival. After freeze-preservation with 40% W/V glycerol at -80 C or with 20% W/V glycerol at -150 C, thawing, washing with sodium chloride solutions, and storage in a sodium chloride-glucose-phosphate solution for 24 hours at 4 C, dog red blood cells had satisfactory recovery values in vitro, acceptable 24-hour post-transfusion survival and long-term survival values, and normal oxygen transport function. Controlled addition and removal of the cryoprotectant, glycerol, helped reduce the amount of osmotic damage to the red blood cells and enhanced freeze-preservation. Osmotic damage can also be prevented by warming the dog blood to a temperature of 22 +/- 2 C prior to centrifugation to concentrate the red blood cells and remove the plasma. This step enhances removal of the cold agglutinins. Another processing step used by the authors was to add a sodium chloride solution to the dog red blood cells before adding the glycerol solution in order to eliminate rouleaux formation.