Platelet viability following storage for 5 days. Influence of holding whole blood for 8 hours at 20 to 24 degrees C before concentrate preparation

Regional blood centers frequently need to hold units of whole blood at 20 to 24 degrees C for several hours after phlebotomy so that sufficient platelet concentrates can be prepared to meet the increasing need. We have evaluated the in vivo viability and in vitro properties of platelets that were prepared from whole blood drawn into citrate- phosphate-dextrose-adenine (CPDA-1) either immediately after phlebotomy or after an 8-hour hold at 20 to 24 degrees C. Platelet concentrates were stored for 5 days at 20 to 24 degrees C in polyolefin containers (PL 732, Fenwal) with end-over-end tumbler agitation. The autologous in vivo recovery (mean +/− SD) and one-half disappearance of 51Cr-labeled platelets prepared immediately after phlebotomy were 44.4 +/− 9.4 percent and 4.0 +/− 0.5 days, respectively. Platelets prepared after the delay of 8 hours showed a recovery of 44.5 +/− 8.4 percent and a one-half disappearance of 4.1 +/− 0.4 days. After 5 days of storage, platelet concentrates showed a mean pH of 7.21 +/− 0.20 when prepared immediately after phlebotomy, and of 7.22 +/− 0.15 when prepared after an 8-hour delay. Mean morphology scores were 280 +/− 33 and 302 +/− 27 for platelets from units prepared immediately after phlebotomy or after a holding period of 8 hours, respectively. Platelets underwent synergistic aggregation after 5 days of storage, independent of the length of time that the units of whole blood were held prior to centrifugation. These studies indicate that platelet concentrates prepared from units of whole blood held initially for 8 hours can be stored for 5 days at 20 to 24 degrees C and survive satisfactorily in vivo and retain in vitro characteristics.     

Characterization of biochemical changes occurring during storage of red cells. Comparative studies with CPD and CPDA-1 anticoagulant- preservative solutions

Citrate-phosphate-dextrose-adenine (CPDA-1), containing 0.25 mM adenine (final concentration) and 25 percent more glucose than citrate-phosphate-dextrose (CPD), has extended the allowable storage time for red cells to 35 days. Studies were conducted to understand better the characteristics of stored CPDA-1 red cells in relation to the properties of stored CPD red cells. Units with hematocrits near 80 percent showed the following: First, adenosine triphosphate (ATP) and total adenine nucleotide levels of red cells stored with CPDA-1 remained essentially constant during the first 3 weeks of storage after which the levels decreased; with red cells stored with CPD, ATP, and adenine nucleotide, levels were decreased even after 1 week of storage. Second, the pattern of the fall in 2,3-diphosphoglycerate was similar in red cells stored with CPD and CPDA-1. Third, changes in plasma and red cell levels of sodium and potassium, and in plasma ammonia levels, were comparable in CPD and CPDA-1 units; changes in cation levels were most pronounced during the initial 2 weeks of storage. Fourth, hemolysis was much greater in units stored in CPDA-1 for 35 days than in units stored in CPD for 21 days. Fifth, residual glucose concentrations were adequate in units drawn in CPDA-1 and stored for 35 days. We conclude that the changes in the biochemical characteristics of units of red cells stored with CPD and CPDA-1 are similar in most instances with the notable exception of the better maintenance of adenosine triphosphate levels in red cells stored with CPDA-1.     

In vivo viability of red blood cells stored in CPDA-2

The marginal viability of erythrocytes stored for 35 days as red blood cell concentrates in citrate-phosphate-dextrose-adenine-one (CPDA-1) was attributed to inadequate nutrient support with adenine and glucose. In an effort to improve the viability of red blood cells following extended storage, a new CPD-adenine and 1.4 times more glucose than CPDA-1. The efficacy of CPDA-2 was evaluated in vivo by measurement of 24-hour postinfusion recovery of 51Cr-labeled erythrocytes which had been stored as whole blood or red blood cell concentrates for 5 to 8 weeks. All red blood cell concentrates, and the whole blood units stored for 35 and 42 days, were held at room temperature for 8 hours prior to processing and/or refrigeration. CPDA-2 yielded significantly higher 51Cr survivals than CPDA-1 and exceeded the accepted criterion for anticoagulant preservative efficacy of 70 percent postinfusion survival of red blood cells after storage for a period of 42 days. Preliminary data supports possible usage to 49 days. Plasma glucose and red blood cell ATP concentration were maintained better in CPDA-2 than in CPDA-1. When compared to historical controls for CPD and CPDA-1 the data suggest that red blood cells stored in CPDA-02 will have superior viability throughout the entire storage period. CPDA-2 is a candidate to replace CPDA-1 as the anticoagulant-preservative solution of choice for red blood cell concentrates.     

Adequate survival of red cells from units "undercollected" in citrate- phosphate-dextrose-adenine-one

The lower limit for the volume of whole blood that may be collected for transfusion into standard blood recipient sets is 405 ml. Each year it is estimated that 82,500 to 161,700 units are drawn which contain between 275 and 405 ml. This study evaluated whether these "undercollected" units would be suitable for transfusion. Twenty normal adults donated both a "standard" unit (450 ml) and an "undercollected" unit (275 ml) in 63 ml of citrate-phosphate-dextrose-adenine-one (CPDA- 1). The units were packed within 4 hours (mean Hct 71%) and stored undisturbed at 4 degrees C for 35 days. Aliquots of 1 to 2 ml of red cells from each unit were then labeled with 51chromium (51Cr) and reinfused into the original donor. The mean 24-hour survival of the 450- ml units was 78.8 percent (SD 12.2, SEM 2.7), while the mean 24-hour survival of the 275-ml units was 87.7 percent (SD 10.7, SEM 2.4; p less than .01). Seven inadvertently undercollected units (mean vol: 295 ml) had a mean 24-hour survival of 91.9 percent. The higher concentration of dextrose and adenine in the undercollected units may improve posttransfusion red cell viability. These data suggest that 275 ml is the minimum acceptable volume for blood donated into CPDA-1.     

The extension of 4 degrees C storage time of frozen-thawed red cells

Blood was drawn from volunteer donors and frozen using the high glycerin, mechanical freezing procedure accepted by the United States Navy. Subsequently, the units of blood were thawed and washed. Various anticoagulants were added, and the red cells were stored in a refrigerator at 4 degrees C for periods of up to 28 days. Chemical analyses were performed periodically. These showed that the addition of the anticoagulants ACD, CPD and CPDA-1 caused the red cells to be preserved better than the currently accepted 0.9-percent NaCl, 0.2-percent glucose solution. In vivo 51Cr viability studies performed on blood stored with CPDA-1 for 14 days showed a 24-hour viability of 78.8 +/- 8.4 percent. In a subsequent study, the blood was stored for 21 days prior to freezing and then was rejuvenated and frozen. The cells were thawed, washed, and stored at 4 degrees C with CPDA-1 for an additional 14 days. The 24-hour viability of these cells was determined to be 74.0 +/- 5.1 percent. These findings show that the postthaw storage time of red cells can be increased greatly over the now-accepted 24 hours, if bacterial sterility can be assured.     

An in vivo comparison of CPD and CPDA-2 preserved platelet concentrates after an 8-hour preprocess hold of whole blood

To see if citrate-phosphate-dextrose-adenine-two (CPDA-2) anticoagulant- preservative had an effect on the viability of platelets, we studied autologous in vivo recovery and survival in humans for platelet concentrates prepared from six units of blood drawn into CPDA-2 and compared them to six units drawn into citrate-phosphate-dextrose (CPD). These units were prepared from whole blood held at room temperature for 8 hours after collection and were then stored for 3 days at 22 ± 2 degrees C. The recovery for platelets preserved in CPD was 39.0 +/0 4.8 percent and for platelets preserved in CPDA-2, 32.5 ± 4.4 percent. The difference was not significant (p greater than 0.10). In order to estimate population differences, in vitro effects on in vivo viability were also evaluated. Six in vitro variables were studied but only pH at 72 hours (r = 0.77), platelet count (r = 0.64), and morphology score (r = 0.66) correlated to recovery. Only pH at 72 hours significantly influenced recovery (p = 0.007). By adjusting for individual pH differences, mean recovery for platelets stored in CPD was 37.5 percent, and for platelets stored in CPDA-2, 34.0 percent. The mean lifespan was 6.7 ± 0.7 days for platelets preserved in CPD and 6.1 ± 1.0 days for those preserved in CPDA-2. Although hemostatic function was not studied, these data support in vitro observations that platelets preserved with CPDA-2 are not different from platelets preserved with CPD, even after 8-hours of storage of whole blood at room temperature prior to platelet concentrate preparation.     

The biochemical effects of CPDA-2-drawn red blood cells of delayed refrigeration prior to component preparation

The effects of an 8-hour hold at 22 degrees C prior to component preparation were evaluated in a split-bag study using nine units of blood preserved in citrate-phosphate-dextrose-adenine (CPDA-2). Each unit was divided in half, platelet-rich plasma removed at 0 or 8 hours, respectively, and the half units of red blood cells stored at 4 degrees C for 42 days. The only red blood cell metabolic differences seen in the bags held 8 hours (compared to those not held) were a 21 percent rise in adenosine triphosphate, which was not significant after 14 days of storage, and a 33 percent loss of 2,3-diphosphoglycerate which resulted in a loss curve similar to that seen with acid-citrate- dextrose blood. The logistic advantages seem to warrant an 8-hour holding period for red blood cells drawn in CPDA-2.     

The in vitro evaluation of modifications in CPD-adenine anticoagulated- preserved blood at various hematocrits

Erythrocytes stored in the new CPD-adenine anticoagulant (CPDA-1) barely met the 70 per cent 24-hour postinfusion 51Cr recoveries on day 35 when stored at hematocrit greater than or equal to 75 per cent. CPDA- 1 differs from CPD in that it has 1.25 times the glucose concentration plus 17.3 mg adenine/63 ml. In an effort to improve the survivability (or viability) of red blood cells following extended storage (35+ days), two new CPD-adenine anticoagulants have been tested in vitro. CPDA-2 and CPDA-3 (both of which contain 34.6 mg/63 ml of anticoagulant or 0.50 mM adenine [final blood concentration], and either 1.75 times or 2.0 times respectively the amount of glucose used in CPD) have been tested for whole blood or red blood cell storage to 42 days. Red blood cell ATP concentrations were better maintained throughout 42 days of storage in both of these formulations than in CPDA-1 at hematocrits that ranged from 40 to 85. Other biochemical parameters (2,3-DPG, pH, plasma hemoglobin) were similar to those of blood stored in CPD or CPDA- 1.     

Posttransfusion survival of red cells frozen for 8 weeks after 42-day liquid storage in AS-3

Current standards recommend that red cells (RBCs) should be frozen within 6 days of donation. There are situations, however, in which it is desirable to freeze RBCs that are older than 6 days, such as for the salvage of rare or autologous units. To determine the therapeutic efficacy of RBCs frozen after prolonged liquid storage, standard units were drawn from nine normal donors and stored at 4 degrees C for 42 days in a nutrient-additive solution, AS-3. 51CrRBC survival assays were performed (24-hour survival: 78.2 +/- 12.4%; n = 8) and the units were frozen at -80 degrees C in glycerol for 8 weeks. After deglycerolization, the mean RBC recovery was 81.0 +/- 4.1 percent and the mean 24-hour 51Cr survival was 78.0 +/- 9.1 percent. The index of therapeutic effectiveness (ITE) was determined by multiplying the postdeglycerolization 24-hour 51Cr survival by the mean RBC recovery (63.3 +/- 9.2). ITE values greater than 60 percent (75% 51Cr survival x 80% RBC recovery) were considered acceptable. Mean adenosine triphosphate levels declined from an initial 3.81 +/- 0.56 micromol per g of hemoglobin to 2.33 +/- 0.55 micromol per g after frozen storage. These findings show that an acceptable percentage of RBCs survives frozen storage after maximum liquid storage (mean ITE greater than 60%). If necessary, RBCs stored in AS-3 can be frozen at any time before 42 days.     

Effect of freezing on the in vivo recovery of irradiated red cells

BACKGROUND: Transfusion-associated graft-versus-host disease can be prevented by gamma radiation of blood components. The increased use of blood components donated for patients by their family members has resulted in an increased demand for the storage and handling of irradiated units, and the ability to freeze the cells would allow storage beyond their current expiration date. STUDY DESIGN AND METHODS: To assess the effect of freezing and deglycerolization on irradiated red cells, studies of autologous radiolabeled red cell recovery were performed using normal volunteers. Each unit of CPDA-1 red cells was immediately divided into two equal volumes. Further handling of each half was identical except that one was irradiated (3500 cGy). The units were grouped under three protocols: I, irradiated on Day 0 and frozen on Day 5 (n = 4); II, irradiated on Day 7, rejuvenated, and frozen on Day 14 (n = 5); and III, irradiated on Day 14, rejuvenated, and frozen on Day 18 (n = 3). All cells were frozen for 3 to 10 months at -80 degrees C. RESULTS: Irradiated and control units showed no significant differences in supernatant potassium or hemoglobin. Autologous 24-hour posttransfusion recoveries (mean +/− SD) for the three groups were: I, 89.7 +/− 5.6 percent (control, 90.6 +/− 3.2%); II, 85.3 +/− 5.7 percent (control, 83.7 +/− 3.0%); and III, 79.5 +/− 1.4 percent (control, 82.6 +/− 5.2%). CONCLUSION: Irradiated red cells can be frozen after being stored under various conditions and can still meet established guidelines requiring 75-percent recovery 24 hours after transfusion.     

The in vivo survival of red blood cells stored in modified CPD with adenine: report of a multi-institutional cooperative effort

In order to provide data in support of licensure applications for citrate-phosphate-dextrose (CPD) supplemented with adenine, a multi- institutional cooperative effort was organized to determine survivability of red blood cells subjected to prolonged liquid storage. Two manufacturers supplied plastic multiple bag blood storage containers prefilled with modified CPD (glucose 25/ greater than the normal concentration) supplemented with adenine (17.0 to 17.3 mg per 63 ml of anticoagulant; 0.25 millimolar approximate final concentration when diluted with 450 ml of whole blood for 35 days showed a mean survival of 80.53 +/- 6.44 per cent (1 SD). Both red blood cell and supernatant plasma biochemical characteristics were comparable to those reported for whole blood stored for 21 days in either acid-citrate- dextrose (ACD) or CPD. Red blood cells from 19 units stored as concentrates for 35 days (Hct 75.03 +/− 3.74%) had a mean survival of 71.38 +/− 10.3 per cent with considerable interdonor variation in survival and interlaboratory variation in some biochemical characteristics. Red blood cells from eight units stored as concentrates (Hct 75.38 +/− 4.30%) for 28 days showed a mean survival of 83.97 +/− 6.10 per cent and biochemical characteristics comparable to those reported for red blood cell concentrates stored in CPD or ACD for 21 days. Modified CPD with adenine as formulated offers an improved anticoagulant for blood banking by extending the permissible red blood cell storage period.     

In vivo viability studies of two additive solutions in the postthaw preservation of red cells held for 3 weeks at 4 degrees C

An optimized additive solution was developed for the postthaw preservation of red cells that contained adenine, glucose, disodium phosphate, and citrate buffer. This solution, called AS-17, was compared to AS-3 solution in a clinical trial using 40 subjects (20 in each arm). Fresh-frozen red cells were thawed and deglycerolized after 1 to 18 months and subjected to a second period of storage in either solution for up to 3 weeks at refrigerator temperatures. Both solutions yielded red cells with 24-hour survivals in excess of 75 percent. Cells stored in AS-3 for 21 days had a mean survival of 77 +/− 8 percent and cells stored in AS-17 a mean survival of 79 +/− 11 percent. The AS-17 solution resulted in improved maintenance of pH, p50, and 2,3 DPG compared to that with AS-3, but both solutions appear adequate for 3 weeks of postthaw storage.     

Storage of saline-adenine-glucose-mannitol-suspended red cells in a new plastic container: polyvinylchloride plasticized with butyryl-n- trihexyl-citrate

Blood collection and component preparation have been performed in integrally connected multiple plastic containers made with a new plastic. This polyvinylchloride (PVC) container plasticized with butyryl-n-trihexyl-citrate (BTHC) is a new material for blood storage; it contains no di(2-ethylhexyl)phthalate (DEHP). After removal of plasma and buffy coat, the red cells were suspended in saline-adenine-glucose-mannitol (SAGM) medium. After 42-day refrigerator storage, the total adenine nucleotide concentration remained the same as the initial concentration in the red cells, whereas ATP levels had decreased to 61 percent of the initial value. The 2,3 DPG concentration was 62 percent of normal on Day 7 and 21 percent on Day 14. Glucose consumption, lactate production, potassium leakage from red cells, and pH levels were similar to those found after storage in DEHP-plasticized containers under the same conditions. After 42 days, hemolysis levels were 0.56 +/- 0.21 percent and 0.42 +/- 0.17 percent in two series of units mixed weekly and 0.70 +/- 0.27 percent in units stored unmixed. Although even higher levels of hemolysis were observed in the units stored unmixed and used for 24-hour posttransfusion survival, the autologous red cell recovery results were excellent (83.2 +/- 5.1%, n = 8). BTHC-plasticized PVC is found to be a suitable material for 42-day storage of red cells in SAGM solution.     

Effect of an 8-hour holding period on in vivo and in vitro properties of red cells and factor VIII content of plasma after collection in a red cell additive system

Extension of the holding time for whole blood units from 6 to 8 hours at ambient temperature should provide enhanced flexibility in the preparation of platelet concentrates (PCs). A paired study was conducted to evaluate the characteristics of stored red cells (RBCs) and plasma prepared from whole blood collected into a red cell additive system (CPD-ADSOL) after an extended holding time. An individual donated a unit of whole blood on two occasions; 1 unit was held for 6 hours before processing and the other for 8 hours. Autologous RBC 24-hour survival levels after 42 days of storage were comparable. Laboratory A, using a 99mTc-51Cr technique, found mean survival levels of 79 percent (6-hour hold) and 78 percent (8-hour hold) (n = 8). Analysis by the single-label procedure found the mean levels to be 82 and 81 percent. Laboratory B, using an albumin 125I-51Cr technique, found mean survival levels of 74 and 72 percent (n = 10). Mean hemolysis and ATP levels were found to be comparable after 42 days of storage following 6- and 8-hour holding periods. 2,3 DPG levels were reduced to a greater degree during the longer hold. The factor VIII levels in plasma frozen for at least a month after 6- and 8-hour holding periods were comparable; thawed plasma contained mean levels of 0.77 and 0.76 units per mL (n = 21). These studies indicate that components prepared by using a CPD-ADSOL system after holding periods of 6 and 8 hours have comparable properties.     

Phosphate ion exchange resin used in the liquid preservation of baboon red cells

Baboon whole blood, collected in 14 percent citrate-phosphate-dextrose anticoagulant solution in plastic bags was stored in 100-ml aliquots at 4 degrees C for 28 days in the presence or absence of 0.75 grams of phosphate anion exchange resin. In vitro and in vivo measurements after autologous transfusions were made to determine whether the phosphate anion exchange resin had any beneficial effect on the blood during storage. The in vitro measurements of red cell 2,3-diphosphoglycerate and P50 were higher throughout the 28 days of storage at 4 degrees C in the blood stored in the phosphate anion exchange resin. After autologous transfusions in six baboons of red cells prepared from whole blood stored at 4 degrees C for 21 days, the 24-hour posttransfusion survival values were 86 +/- 6 percent (mean +/- SD) in the presence of resin and 83 +/- 6 percent in the absence of resin. In five other baboons, red cells prepared from 28-day-old blood showed a mean 24-hour posttransfusion survival of 82 +/- 4 percent in the presence of resin and 75 +/- 4 percent in the absence of resin. The addition of a phosphate anion exchange resin to the citrate-phosphate-dextrose anticoagulant provided better maintenance of red cell 2,3-diphosphoglycerate concentrations and P50 levels during storage of whole blood at 4 degrees C, and red cells prepared from whole blood stored in this solution had better oxygen transport function than red cells prepared from blood stored without resin. Red cell adenosine triphosphate concentrations and 24-hour posttransfusion survival values were similar whether or not the anticoagulant contained resin.     

Effects of 4000 rad irradiation on the in vitro storage properties of packed red cells

Immunosuppressed patients who require red cell transfusions receive irradiated (1500-3000 rad) packed red cells. These cells are irradiated immediately before infusion. If a large group of patients become immunosuppressed due to exposure to radiation or chemicals, the ability to supply large volumes of irradiated blood at the time of use might not be possible. An alternate solution to providing quantities of irradiated blood is to irradiate the units prior to storage. This study presents in vitro data comparing storage of paired packed red cell units either irradiated or not irradiated. Five units of fresh blood drawn into citrate-phosphate-dextrose-adenine (CPDA-1) were packed to a hematocrit of 75 +/- 1 percent, and then each unit was divided in two equal parts. One of each pair was irradiated (4000 rads), and both parts of each unit were stored for 35 days at 4 degrees C. Samples were analyzed every 7 days. Irradiation caused a slight drop in red cell adenosine triphosphate and 2,3 diphosphoglycerate and a slight increase in plasma hemoglobin compared to controls. Methemoglobin, pH, and glucose consumption were identical to the controls. The evidence indicates that irradiation did not cause biochemical or metabolic changes in the red cells that would lead us to suspect a difference between irradiated and nonirradiated stored red cells in function or viability. These negative findings require in vivo confirmation.     

Red cell storage for 56 days in modified DPD-adenine. An in vitro evaluation

The modified CPD-adenine anticoagulants CPDA-2 and CPDA-3 were developed to improve red blood cell storage to 35 days, since CPDA-1 was found marginal at 35 days in high hematocrit samples. In this study red blood cell storage was extended to 56 days. In vitro correlates of viability were monitored to determine the feasibility of evaluating in vivo the ability of CPDA-2 and CPDA-3 to extend storage past 35 days. The data suggest that red blood cells stored up to 56 days may have acceptable viability, providing the possibility of extended storage for the military and certain special civilian situations.     

Effect of delayed refrigeration on plasma factors in whole blood collected in CPDA-2

Extension of the time within which whole blood may be separated into components offers logistic advantages for the operation of remote mobile drawing teams. We evaluated the effect of an 8-hour hold of whole blood at room temperature before preparation of components. Plasma coagulation activity and opsonic factor content were studied in 14 units drawn into the anticoagulant-preservative solution citrate-phosphate-dextrose-adenine (CPDA-2). At the time of collection, an additional 7-ml aliquot was drawn into 1 ml of CPDA-2, the plasma separated and frozen immediately. Components were prepared from whole blood units allowed to rest undisturbed at 22 +/- 1 degrees C for 8 hours. After 8 hours, a significant decrement of about 10 percent was found in the concentration of fibrinogen, plasminogen, fibronectin, and activity of Factor V. Factor VIII activities (VIIIAHF and VIIIAGN) were not significantly different after 8 hours. Our results indicate that room temperature storage for 8 hours before component processing has minimal effects on potentially labile plasma protein factors using CPDA-2 anticoagulant-preservative solution.     

Multiple-unit and second transfusions of red cells enzymatically converted from group B to group O: report on the end of phase 1 trials

BACKGROUND: It has previously been shown that 1 and 2 units (200 - 400 mL) of red cells (RBCs) enzymatically converted from group B to group O by treatment with alpha-galactosidase (ECO RBCs) are safe and efficacious when transfused to normal group O or A persons. STUDY DESIGN AND METHODS: The current report describes studies in which 1) normal group A and O subjects received large volumes of these cells (3 units), 2) some group O subjects underwent transfusion several months later, and 3) ECO RBCs were prepared by the use of recombinant coffee bean alpha-galactosidase and transfused to a group O subject, to demonstrate the in vivo equivalence of ECO RBCs, whether prepared with native or recombinant alpha-galactosidase. RESULTS: Clinical evaluation (hematologic tests, chemistry analysis, urinalysis) and serologic analyses did not reveal any evidence of subtle or acute transfusion reaction or significant increase in preexisting anti-B titer. ECO RBC survival within the circulation of the recipients was normal (24-hour survival, 95.5 +/− 0.9%; t1/2, 34.7 +/− 6.1 days; n = 8 transfusions), and the efficacy of the transfusions was manifested in elevations in recipient hemoglobin and hematocrit (hemoglobin increase, 1.5 +/− 0.6 g/dL; hematocrit increase, 3.6 +/− 1.6%; n = 8 transfusions). CONCLUSION: ECO RBCs are safe and efficacious when transfused more than once or in multiple-unit volumes to group O or A subjects, and ECO RBCs prepared with recombinant or native enzyme are equivalent in vivo.     

Posttransfusion survival (24-hour) and hemolysis of previously frozen, deglycerolized RBCs after storage at 4 degrees C for up to 14 days in sodium chloride alone or sodium chloride supplemented with additive solutions

BACKGROUND: Previously frozen human RBCs currently are glycerolized and deglycerolized by the use of open systems that limit storage of the deglycerolized RBCs at 4 degrees C to only 24 hours. STUDY DESIGN AND METHODS: Healthy male volunteers who met AABB requirements for blood donors (n = 38) were studied. A volume of 450 mL of blood was collected into CPDA-1. The RBC concentrates were stored at 4 degrees C for 3 to 6 days before being frozen with 40-percent (wt/vol) glycerol and stored at -80 degrees C. The RBCs were deglycerolized, resuspended in 0.9-percent sodium chloride and 0.2-percent glucose (SG) solution or SG solution supplemented with AS-1, AS-3, or AS-5, and stored in the resuspension medium at 4 degrees C for 14 days. RESULTS: The mean +/- SD freeze-thaw-wash process recovery was 90.0 +/- 4.0 percent for all 38 units. The mean 24-hour posttransfusion survival value was 79 percent for deglycerolized RBC stored at 4 degrees C for 7 days in SG alone, SG plus AS-3, or SG plus AS-5. Deglycerolized RBC that were stored at 4 C for 14 days in SG supplemented with AS-1, AS-3, or AS-5 had a mean 24-hour posttransfusion survival of 74 percent. After 7 days of storage of deglycerolized RBCs in SG alone, the mean hemolysis was 3. 7 percent. After 14 days of storage of deglycerolized RBCs in SG supplemented with AS-1, AS-3, or AS-5, the mean hemolysis was 2.5 percent. CONCLUSIONS: The levels of hemolysis did not correlate with the 24-hour posttransfusion survival values.