Human red cells frozen by various methods have been stored in the frozen state at -80 degrees C for as long as 21 years. This report discusses: red cells frozen with 42 percent weight per volume (wt/vol) glycerol in an ionic medium in a polyvinylchloride (PVC) plastic bag using the Cohn method; red cells frozen with 45 percent wt/vol glycerol in a low ionic medium in a PVC plastic bag using the Huggins method; red cells frozen with 40 percent wt/vol glycerol in an ionic medium in a polyolefin plastic bag using the Meryman-Hornblower method; and red cells frozen with 40 percent wt/vol glycerol in an ionic medium in a standard 600-ml or an elongated 800-ml PVC plastic primary collection bag with an adapter port using the Naval Blood Research Laboratory (NBRL) method. After frozen storage for as long as 21 years by the four methods described above, the thawed red cells were deglycerolized with 50 to 150 ml of 12 percent sodium chloride and 1.5 to 2.0 l of sodium chloride-glucose or sodium chloride-glucose-phosphate solution. After washing and storage at 4 degrees C for 24 hour, the red cells had a mean freeze-thaw-wash recovery value of 90 percent, a mean 24-hour posttransfusion survival value of 85 percent, a mean index of therapeutic effectiveness of 75 percent, normal or slightly impaired oxygen transport function, and minimal hemolysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Repeated intravenous infusions of radioiodinated human albumin and human fibrinogen into normal healthy baboons over a six- to twelve-month period were used for measurements of plasma volume. The radioiodinated fibrinogen gave values for plasma volumes that were quite variable and 20-25% lower than those obtained using radioiodinated albumin. During this period of repeated infusions, there was no apparent reduction in the survival of the human protein in the baboon circulation that would indicate immunization had occurred. Radioiodinated baboon fibrinogen and human fibrinogen administered to the same baboon showed similar survival patterns.

A method has been developed for separation of erythrocytes on the basis of size using counterflow centrifugation. Human red blood cells with an original mean corpuscular volume (MCV) of 89.2 +/- 4.1 fl were isolated, free of plasma proteins and other cell contaminants, into seven fractions ranging in size from 77.0 +/- 2.7 fl to 98.5 +/- 4.8 fl. The ratio of the age-related enzyme, erythrocyte glutamic oxaloacetic transferase (EGOT), to hemoglobin (Hb) increased progressively through the fractions, suggesting a correlation between erythrocyte volume and age. Reticulocytes, though present in all fractions, were selectively enriched in the larger subpopulations. To verify the biochemical evidence that erythrocytes decrease in volume with aging, in vivo cohort labeling of red blood cells with 59Fe was performed in baboons. A similar relationship of EGOT to Hb was observed to that in the human subpopulations. The peak activity of 59Fe/RBC appeared initially in the red blood cells with the highest MCV and progressed from the erythrocytes with the largest MCV to the erythrocytes with the smallest MCV over the next 10-12 weeks, confirming the hypothesis that red blood cells decrease in volume as they age. The technique of counterflow centrifugation appears to provide a simple, rapid, and reproducible method for the separation of erythrocytes on the basis of size.