Incorporation of Acetate-2-¹⁴C into Human Erythrocyte Stroma as a Function of Storage

Abstract

IN an effort to elucidate the changes which may occur in the dynamic state of stroma constituents during storage of human erythrocytes (reported in part at the “Symposium on the Structural and Cellular Dynamics of the Red Blood Cell”, June 1953: National Research Council, Washington, D.C.), we have studied the incorporation of acetate-2-¹⁴C into red cell stroma under in vitro conditions using a technique previously described¹. Venous blood from healthy donors was drawn under sterile conditions into cold ACD solution (each 100 c.c. of the ACD solution contained 2.45 gm. glucose, 1.37 gm. sodium citrate and 0.50 gm. of citric acid; 0.24 c.c. of ACD solution was added to each c.c. of blood) and then distributed as aliquots into an appropriate number of sterile tubes. These tubes were carefully sealed to prevent evaporation and were stored in two groups, one group at 4° C. and the other group at 37° C. At the intervals indicated in Fig. 1, tubes were removed from the storage conditions and the incorporation of acetate-2-¹⁴C into the total stromal fraction of the stored cells was determined. The composition of the incubation mixture for measurement of acetate incorporation was: 12.0 ml. of ACD-preserved whole blood, 1.5 ml. isotonic phosphate buffer at pH 7.4, 0.6 ml. of 0.3 M glucose in 0.9 per cent sodium chloride, and 1.5 ml. of sodium acetate-¹⁴C (approximately 80 mgm.) in 0.9 per cent sodium chloride. The total amount of radioactivity added as labelled acetate varied from 4.9 to 6.3 × 10⁶ disintegrations per minute per 12 ml. of whole, preserved blood.