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Blood refrigerated in acid-citrate-dextrose is known to undergo marked derangement in carbohydrate metabolism and loss of viability during storage. Previous studies of the "storage lesion" have indicated that this metabolic deterioration is arrested once the red cell is introduced into the active circulation of the recipient, and that the ability of the red cells to withstand further in vitro storage is regained (1).The addition of adenosine to stored blood has been shown to produce a repletion of cellular organic phosphates and an increased ability of the cells to oxidize glucose aerobically (2). Further data will be presented in this paper on the reversal of other storage changes and in particular the improved maintenance of the post-transfusion viability of the erythrocyte. MATERIALS AND METHODS Blood

The progressive morphological and biochemical alterations which develop in the erythrocyte during in vitro storage are not the result of cell aging' in a physiological sense (2), nor do they appear to be induced by harmful substances in the extra-erythrocyte environment (3). Rather, the "storage lesion" is considered to be a primary metabolic failure of the in vitro erythrocyte. The various measurable abnormalities are regarded only as indicators of this metabolic lesion, since no single abnormality in itself has been shown to be critical to cell viability.Since the red cell derives its energy primarily from carbohydrate metabolism, the glycolytic scheme and associated high energy phosphate compounds would seem important in a consideration of an abnormality which progresses to ultimate loss of cell viability. Furthermore, it has been shown that characteristic changes appear in the glycolytic structure of the erythrocyte during storage (4) which can be correlated with the posttransfusion survival of blood. In this study of the reversibility of the storage lesion, the phosphate partition, particularly adenosine polyphosphates, was employed as an indicator of the metabolic potential of the cell. The significance of these measurements was evaluated by the posttransfusion survival of the stored erythrocytes. MATERIALS AND METHODSRabbit blood was collected by intracardiac puncture using sterile precautions and stored at 4°C . in ACD (acid-citrate-dextrose) as previously described (2). artery. The recipient cell mass had been tagged previously by the injection of fresh rabbit erythrocytes containing labeled hemoglobin (Fe59). Thus the exact amount of exchange of the recipient's erythrocytes with stored blood was calculated from the decrease in circulating radioactivity immediately after the exchange transfusion. The subsequent destruction of stored blood over the following 12 hours was calculated from the increase in the specific activity of erythrocytes since only the nonradioactive transfused cells were destroyed at an increased rate. With this type of short-term experiment in animals, the reversal of the biochemical changes in the stored erythrocytes was investigated.In man, studies were undertaken to determine the ability of an erythrocyte, after an initial period of storage and transfusion, to withstand a second period of in titro storage. Patient 1 (H. S., Type A) with aregenerative anemia was transfused repeatedly over a period of three weeks with Type 0 blood which had been in storage for three weeks. Blood was then withdrawn from H. S. and stored for three weeks before transfusion into a Type A recipient. The survival of the Type 0 cells was determined in the second recipient by the differential agglutination technique. Patient 2 (K. K., Type 0) with aregenerative anemia was given 20 units of three-week stored Type 0 cells over a period of 10 weeks. At the completion of these transfusions, when it was almost certain that all of his circulatory erythrocytes were composed of red cells which had undergone three w...

Erythrocytes stored in an acid-citrate-dextrose preservative (ACD) at 4 to 7°C. undergo morphological and biochemical alterations. The cell becomes more sphere-shaped with an associated increase in osmotic and mechanical fragilities, the glycolytic rate and organic phosphate compounds are decreased, inorganic phosphate is increased, and the differential between cell potassium and sodium is decreased (1). These altered cells, when transfused to a recipient, are destroyed in an amount dependent upon the duration of storage of the blood. Thus, storage for three weeks in ACD results in a viability 2 of about 85 per cent and for four weeks in a viability of about 60 per cent (2, 3).Since the normal human erythrocyte population is destroyed in vivo at a rate of 0.8 to 1.0 per cent per day, the survival of erythrocytes after three to four weeks of storage is consistent with the expected in vivo loss from aging. The present study was undertaken to determine whether the changes of storage were related to a normal or accelerated aging process, or whether they represented cell damage due to in vitro storage conditions.

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