2,3diphoephoglyceric acid appears to be an important regulator of the oxygen dissociation curve of hemoglobin in intact red blood cells. The rate of loss of 23-DPG under various storage conditions therefore was investigated. ZB-DPG disappeared rapidly from conventional preservative media, CPD, and ACD solutions. After only two weeks' storage, 65 per cent to 85 per cent of erythrocyte 23-DPG had been lost from ACD-stored blood and slightly less from CPD-stored blood. Although the addition of adenine to ACD solution aided in the maintenance of ATP levels, it hastened the rate of loss of 23-DPG. The rate of 2,3-DPG depletion was strongly dependent on pH. In more alkaline storage media, levels of this compound were relatively well maintained for as long as two, or even three weeks. However, under these circumstances ATP maintenance was less satisfactory. The levels of 23-DPG and ATP in red blood cells incubated in fresh plasma at 37C, pH 7.4, to simulate the conditions after reinfusion of stored celb also was investigated. ATP levels remained relatively stable under these circumstances and 2,3-DPG levels were restored gradually. However, the repletion of 28-DPG was sufficiently slow so that even after eight hours only approximately one third of the 23-DPG which had been lost was regenerated. Thus, stored blood may fail to transport oxygen efficiently for many houra after reinfusion.
Summary. The addition of adenine and ascorbate to citrate‐phosphate‐dextrose solution has been shown to prolong the maintenance of 2,3‐DPG levels of stored blood. The final concentrations of adenine and sodium ascorbate in the blood‐preservative mixture were 0.5 and 5.05 mm, respectively. In most cases, red cells stored in this preservative maintained levels of 2,3‐DPG greater than 50% of fresh values for 28 days. Red‐cell viability was not adversely affected by ascorbate. The mechanism by which ascorbate produced this effect is unknown.
The storage of red blood cells (RBC) for extended periods in artificial media without plasma has been studied. Blood was collected in either heparin or ACD solution; the plasma was removed; and one or two volumes of a solution containing 2 to 3 mM adenine, 5 to 60 mM Na2HPO4 55 mM glucose, and 120 to 140 mM NaCl was added to the packed RBC. Control samples were stored in ACD plasma containing an equivalent concentration of adenine (ACD‐ad). Viability studies were done on three consecutive days in each of 22 subjects, using the subject's own blood stored in various preservatives for 41 to 57 days. After this period of storage, the mean viability of ACD‐ad stored blood was 73.5 per cent and of erythrocytes stored in artificial media, 74.4 per cent. Cells stored for longer periods had diminished viability, but the viability of cells stored in artificial media was equivalent to or superior to that of controls. After 42 days' storage, 2,3 DPG levels of RBC stored in artificial media were higher than those of controls, and in some instances, the 2,3 DPG content was one‐third to two‐thirds that of fresh blood. Some of the potential advantages of this system for blood preservation are: 1. The suspending medium is discarded prior to infusion so that less potentially toxic substances are administered. 2. Plasma is removed at the beginning of storage so that labile factors are available for fractionation. 3. 2,3 DPG levels are higher, so that, theoretically, the oxygen‐delivering capacity of the transfused cells is greater.
Blood from six donors was stored in two experimental blood preservatives, CPD-adenine-sodium ascorbate and bicarbonate-adenine-glucose-phosphate-mannitol. Control aliquots were stored in CPD-adenine and ACD-adenine preservatives. Half of each donor’s blood was stored without mixing and half was mixed vigororously 5 days each week. Glucose, ATP, and 2,3-DPG levels were measured at various times during storage in both the mixed and unmixed bags. Mixed bags maintained higher levels of glucose, ATP, and 2,3-DPG throughout storage. Frequent mixing was shown to be essential for the maintenance of 2,3-DPG with these preservatives.
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