Blood components produced from whole blood using the Atreus processing system

BACKGROUND: The Atreus 2C+ system (Gambro BCT) automates whole blood (WB) processing into a single device. This study compared the quality of red blood cells (RBCs), fresh‐frozen plasma (FFP), and buffy coats (BCs) made from WB held with or without active cooling. STUDY DESIGN AND METHODS: WB was collected into Atreus disposables and stored with (n = 20) or without (n = 20) active cooling for 14 to 18 hours at 22 ± 2°C before processing with the Atreus. Two RBC leukodepletion filters were assessed, and markers of RBC quality were tested to Day 42. BCs were held for 3 hours before testing, plasma was tested, and samples were frozen for coagulation analysis. RESULTS: RBCs met UK specifications for volume, hemoglobin content (48 ± 5 g), and hematocrit (Hct). Hemolysis, adenosine triphosphate, 2,3‐diphosphoglycerate, potassium, glucose, and lactate throughout storage were all within expected ranges. No differences were seen in RBC produced from WB held with or without active cooling. FFP units met UK specification for volume, total protein, cellular contamination, and coagulation factors. No differences were seen in FFP produced from WB held with or without active cooling. The Hct of BCs produced from WB held without active cooling was lower than in BCs from WB held with active cooling; no differences in activation were seen. CONCLUSION: From these in vitro data, blood components produced using the Atreus appear suitable for clinical use, with no clinically significant difference in the quality of components from WB held at ambient temperature overnight with or without active cooling.     

Stability of coagulation factors in plasma prepared after a 24‐hour room temperature hold

BACKGROUND: The manufacture of fresh‐frozen plasma (FFP) requires that plasma be frozen within 8 hours of collection and 24‐hour frozen plasma requires 1 to 6°C refrigeration before freezing. Manufacture of plasma after a room temperature hold for 24 hours, while convenient, could compromise clotting factor levels. STUDY DESIGN AND METHODS: Pairs of FFP and 24‐hour room temperature–frozen plasma (PLT‐rich plasma [PRP]‐24HRTFP) were manufactured from PRP after a room temperature hold for 8 and 24 hours, respectively. Additional whole blood (WB) donations were kept at room temperature for 24 hours before plasma manufacture (WB‐24HRTFP). The frozen plasma products were stored at −18°C, thawed, and then stored at 1 to 6°C, with coagulation factor assays performed for up to 7 days. RESULTS: On the day of thaw, Factor (F)VIII was lower in PRP‐24HRTFP by 13% (p = 0.002) but not in WB‐24HRTFP (p = 0.3) compared to FFP. All other clotting factors were within normal range. During the postthaw period FVIII and FV declined 25 and 6%, respectively, in WB‐24HRTFP and 23 to 50% in the paired products; however, the difference between both types of 24HRTFP and FFP is insignificant by Day 7 (p > 0.05). Other clotting factors either were unchanged or showed minimal reduction (<15%). CONCLUSION: Plasma manufactured after a 24‐hour room temperature hold contains coagulation factors comparable to FFP except for a possible reduction of up to 20% in FVIII. This plasma appears suitable as a transfusable product and extension of liquid storage to 7 days merits consideration.     

The quality of fresh-frozen plasma produced from whole blood stored at 4 degrees C overnight

BACKGROUND: The aim of this study was to assess whether the quality of FFP produced from whole blood stored at 4 degrees C overnight is adequate for its intended purpose. STUDY DESIGN AND METHODS: Fresh-frozen plasma (FFP) separated from whole blood (n = 60) leukodepleted (LD) after storage at 4 degrees C overnight (18-24 hr from donation, Day 1 FFP) was compared with that LD within 8 hours of donation (Day 0 FFP, the current standard method). RESULTS: In more than 95 percent of Day 1 FFP units, levels of factor (F) II, FV, FVII, FVIII, F IX, FX, FXI, and FXII were greater than 0.50 U per mL except for von Willebrand factor (VWF) antigen and FVIII, where 92 and 87 percent of units, respectively, contained greater than 0.50 IU per mL. Compared with historical data on FFP stored for 8 hours, fibrinogen, FV, FVIII, and FXI were reduced by 12, 15, 23, and 7 percent, respectively, but other factors were not significantly reduced. Levels of VWF-cleaving protease activity were not different between FFP prepared from paired units of blood (n = 3) held for 8 or 24 hours, but were below the reference range in an additional 2 of 6 units held for 24 hours. The activities of protein S, protein C, antithrombin III, and alpha(2)-antiplasmin were reduced by less than 10 percent in Day 1 FFP (n = 20), but with final levels above the lower limit of the normal range in greater than 95 percent of units. Activated FXII antigen was not significantly raised in plasma stored for 18 to 24 hours, but levels of prothrombin fragment 1 + 2 were slightly increased (0.88 ng/mL, 18-24 hr; 0.65 ng/mL, < 8 hr). CONCLUSION: These data suggest that there is good retention of relevant coagulation factor activity in plasma produced from whole blood stored at 4 degrees C for 18 to 24 hours and that this would be an acceptable product for most patients requiring FFP.     

Improved maintenance of 2,3 DPG and ATP in RBCs stored in a modified additive solution

BACKGROUND : All currently used systems for the storage of RBCs result in loss of 2,3 DPG and an associated increase in affinity for oxygen. Previously, it was demonstrated that a hypotonic additive solution for RBC storage (Erythro‐Sol) ^{4 , 5} resulted in prolonged maintenance of 2,3 DPG when blood was collected in 0.5 CPD (half‐strength CPD), but not when full‐strength CPD was used. The present study aims at improving the quality of stored RBCs collected in ordinary CPD. STUDY DESIGN AND METHODS : A new formulation of Erythro‐Sol (Erythro‐Sol 2) (pH 8.8) in a larger volume (150 mL) was compared with Erythro‐Sol (Erythro‐Sol 1). In vitro measures during 49 days of storage in the two additives were compared using WBC‐depleted RBCs after whole‐blood collection in CPD and separation in an automated blood separation instrument (Optipress II, Baxter Healthcare). RESULTS : The maintenance of RBC ATP and 2,3 DPG was significantly better in Erythro‐Sol 2 than in Erythro‐Sol 1. The ATP concentration rose to approximately 30 percent above initial level in both systems; however, the maximum occurred on Day 21 in Erythro‐Sol 2 as compared with Day 14 in Erythro‐Sol 1. In RBCs stored in Erythro‐Sol 2, the mean RBC 2,3 DPG concentration increased to 14 percent above initial level on Day 7, then decreased to the initial level on Day 14, whereas in Erythro‐Sol 1, the 2,3 DPG had decreased to 85 and 50 percent on Days 7 and 14, respectively. Both intracellular pH and extracellular pH were slightly higher in Erythro‐Sol 2 than in Erythro‐Sol 1 units but decreased rapidly during the first storage week, which seems to have been the major reason for the limitation in the time of maintenance of 2,3 DPG. Hemolysis was very low in both systems, 0.14 to 0.17 percent on Day 49. The additional amount of inorganic phosphate submitted with Erythro‐Sol 2 did not raise concern because the phosphate content in the storage medium, being 1.3 ± 0.2 mmoL on Day 0, decreased to values below 1 mmoL during most of subsequent storage. CONCLUSION : Erythro‐Sol 2 is an improved additive solution for the storage of RBCs.     

Active cooling of whole blood to room temperature improves blood component quality

BACKGROUND: Many countries use cooling plates to actively cool collected whole blood (WB) to room temperature. Until now, no paired comparison had been performed, and it was our aim to compare the effect of active versus no active cooling on the in vitro quality of WB and subsequently prepared blood components. STUDY DESIGN AND METHODS: Two units of WB were pooled and divided shortly after donation. One unit was placed under a butane‐1,4‐diol plate to obtain active cooling; the other was placed in an insulated box with other warm units to mimic worst‐case holding conditions. WB was held overnight and processed into a white blood cell (WBC)‐reduced red blood cells (RBCs), buffy coat (BC), and plasma. The BCs were further processed into platelet (PLT) concentrates. RBCs were stored for 42 days, and PLT concentrates for 8 days (n = 12 paired experiments). RESULTS: After overnight storage, ATP content of the RBCs was 4.9 ± 0.3 µmol/g Hb for actively cooled WB versus 4.5 ± 0.4 µmol/g Hb for not actively cooled WB (p < 0.001). On Day 42 of storage, RBCs prepared from this WB contained 3.1 ± 0.3 µmol ATP/g Hb with active cooling versus 2.6 ± 0.3 µmol/g Hb without (p < 0.001). Hemolysis on Day 42 was 0.35 ± 0.08% with active cooling and 0.67 ± 0.21% without (p < 0.001). No effect was observed on the in vitro quality of plasma, BC, or PLT concentrates. CONCLUSIONS: Active cooling of WB results in improved ATP levels and less hemolysis in WBC‐reduced RBCs, although the clinical implications are unclear. It has no effect on the in vitro quality of plasma or PLT concentrates.     

Characterization of blood components separated from donated whole blood after an overnight holding at room temperature with the buffy coat method

BACKGROUND: With buffy coat (BC) processing of whole blood (WB) donations, increase in WB storage time to facilitate overnight holding before the separation of blood components would be a logistically attractive development. This study undertakes a comparative in vitro characterization of blood components prepared from WB samples that were either processed within 8 hours or stored overnight at room temperature before processing by the BC method. STUDY DESIGN AND METHODS: The WB units (400 mL) collected were either processed within 8 hours (fresh blood) or stored overnight (overnight blood) at room temperature. WB units were separated into individual‐component red blood cells (RBCs), BC, and plasma. The in vitro quality of these blood components (RBCs, pooled platelet concentrates [PCs], and plasma) was analyzed during storage. RESULTS: Levels of 2,3‐diphosphoglycerate (2,3‐DPG) were found to be significantly lower immediately after processing, compared with the fresh WB samples, in RBCs that had been separated from an overnight‐hold sample. However, this difference was not apparent after 14 days of storage. In pooled PCs, measurements for glucose, lactate, PO₂, PCO₂, extent of shape change, and hypotonic shock response were similar. The platelet yield in PCs prepared from an overnight‐hold WB sample was significantly higher, while CD62P expression and annexin V binding were lower (p < 0.05). For frozen plasma (FP), no significant differences were observed for the coagulation factors (F)II, FVII, FV, F IX, FX, and FXI; fibrinogen; and von Willebrand factor content between the 8‐ and 24‐hour FP. The FVIII was the component that was most sensitive to the prolongation of production time and it only had 80% of the activity of the 8‐hour FP. CONCLUSION: These data suggest that blood components (RBCs, pooled PCs, and FP) separated from WB that has been stored overnight at room temperature by the BC method are of acceptable quality.     

Storage of whole blood before separation: the effect of temperature on red cell 2,3 DPG and the accumulation of lactate

BACKGROUND: Although whole blood intended for component preparation is commonly left to cool at ambient temperature, knowledge is insufficient as to the effects this may have on red cell quality, in particular after a prolonged hold.
STUDY DESIGN AND METHODS: Whole blood collected in ACD-A (7% wt/wt) and CPD (12% wt/wt) was incubated at 4, 10, 15, 20, 25, and 30°C for 24 hours. Blood gases, pH, bicarbonate, glucose, lactate, and red cell 2,3 DPG were investigated.
RESULTS: When the blood was stored at 30°C, the 2,3 DPG concentration decreased within 4 hours from 858 ± 106 to 316 ± 172 mmol per mol of hemoglobin (a 63% decrease); 99 percent was lost within 18 hours. At 25°C, 46 percent was lost within 4 hours and 94 percent within 18 hours; at 20°C, the decrease at 18 hours was 62 percent and that at 15°C was 24 percent. No loss of 2,3 DPG was observed at 4°C and 10°C storage. No difference was attributable to the anticoagulant used. After 24 hours, the lactate concentration at 15°C was 2.9 times the original, that at 20°C was 3.8 times the original, that at 25°C was 7.0 times, and that at 30°C was 9.2 times.
CONCLUSIONS: With current anticoagulants, storage of whole blood at temperatures of 25 to 30°C before separation causes a great and rapid loss of 2,3 DPG and an accumulation of acid metabolites. In a hold of blood for >4 hours, rapid cooling is desirable to avoid initial loss of 2,3 DPG.     

Double red cell concentrates –in vitro quality after delayed refrigeration

Automated collection of red cell concentrates (RCC) presents a number of potential advantages to donors, blood services and recipients, and allows the collection of finished components from sites that are remote from a blood centre. However, data are lacking on how long the collected RCC may be stored at ambient temperature prior to their final storage at 4 °C. In this study, the Haemonetics Cymbal device was used to collect RCC using citrate, phosphate and dextrose (CPD‐50) anticoagulant. A total of 10 procedures each yielded two leucodepleted RCC in saline, adenine, glucose and mannitol (SAGM) additive solution. One of each pair of RCC was kept warm in an insulated transport bag for 8 h and the other for 6 h. In vitro assessments of the quality of the RCC were made during subsequent 42‐day storage of the RCC at 2–6 °C, and compared with reference data. All collected RCC were within UK and European limits for volume, haematocrit and haemoglobin content. Haemolysis was within specification at Day 42 and was no different in RCC held warm for 6 or 8 hours, but tended to be higher than reference data from whole blood derived RCC. ATP, 2,3 DPG and supernatant potassium levels were all similar in RCC held warm for 6 or 8 hours and reference data. We conclude that the Cymbal device may be used to collect two RCC in SAGM, and the in vitro assessment indicates that RCC may be stored without refrigeration for up to 8 h following collection, prior to final storage at 4 °C.     

Rejuvenation capacity of red blood cells in additive solutions over long-term storage

BACKGROUND: Red blood cells (RBCs) are Food and Drug Administration (FDA)‐approved for 42‐day storage with the use of additive solutions (ASs). However, adenosine triphosphate (ATP) and 2,3‐diphosphoglycerate (2,3‐DPG) levels in the RBCs decline over this time. These constituents may be restored by treatment with rejuvenation (REJ) solutions. This study was done to assess the response capability of RBCs from 30 to 120 days of storage in three FDA‐licensed RBC storage solutions after incubation with a rejuvenating solution of pyruvate, inosine, phosphate, and adenine. STUDY DESIGN AND METHODS: Three units each of RBCs in approved AS (AS‐1 [Adsol, Fenwal, Inc.], AS‐3 [Nutricel, Medsep Corp.], and AS‐5 [Optisol, Terumo Corp.]) were stored under standard conditions at 1 to 6°C for up to 120 days. Aliquots (4 mL) on Days 30, 42, 60, 80, 100, and 120 (±2 days) were REJ by incubating with Rejuvesol (Encyte Corp.). Control untreated and REJ aliquots were extracted using perchloric acid and stored at ?80°C until assayed for 2,3‐DPG and ATP. RESULTS: RBCs responded to REJ by increasing DPG and ATP contents. The response declined linearly at 0.070 ± 0.008 µmol DPG/g hemoglobin (Hb)/day and 0.035 ± 0.004 µmol ATP/g Hb/day with no differences between ASs. CONCLUSION: We conclude that Rejuvesol is able to restore ATP and 2,3‐DPG levels in RBCs stored up to 120 days in AS. The response diminishes as storage time increases. This rejuvenation (REJ) capability does not seem useful for routine assessment of RBC anabolic capacity in research programs, but may be useful to the investigator when studying unique and novel treatment methods.     

Coagulation profile of liquid‐state plasma

BACKGROUND: Use of liquid plasma (LP) has been reported as early as the mid 1930s. Unlike fresh‐frozen plasma (FFP), LP is maintained at 1 to 6°C for up to 40 days after collection and processing. Despite its approved use by the US Food and Drug Administration, the coagulation profile of LP is incompletely described. In this study we evaluate the coagulation profile of LP stored up to 30 days. STUDY DESIGN AND METHODS: LP was prepared by removing plasma from nonleukoreduced whole blood within 24 hours of collection. Three LP units from each ABO group were collected and stored at 1 to 6°C. Plasma aliquots were obtained at Postcollection Days 1 to 5, 10, 15, 20, 25, and 30 and then stored at ?70°C. Each aliquot was tested for prothrombin time, activated partial thromboplastin time, and other coagulation and fibrinolytic factors. RESULTS: There was a significant decrease in Factor (F)V, FVII, FVIII, von Willebrand factor (VWF), protein S (PS) activity, and endogenous thrombin potential on Day 15 compared with Day 1. No significant difference was observed for PS antigen, D‐dimer, or thrombin‐antithrombin complex. At least 50% activity of all measured factors was noted on Day 15, compared to Day 1. Considerable heterogeneity was observed between the different blood groups for FVII, FVIII, and VWF. CONCLUSION: These data demonstrate that LP maintains at least 50% of factor activity and thrombin‐generating capacity up to 15 days of refrigerated storage. It may be more appropriate to limit LP storage and supplement with FFP when used for management of massively bleeding patients.     

Overnight storage of whole blood: a comparison of two designs of butane-1,4-diol cooling plates

BACKGROUND: Whole blood (WB) can be stored overnight before processing, provided that it is quickly cooled to room temperature (20-25 degrees C), for example, with butane-1,4-diol plates. A new design of cooling plates became available (CompoCool-WB, Fresenius HemoCare), where WB must be placed vertically against the plates, versus placing of WB under plates in the current version (Compocool). This study compared cooling efficiency and in vitro quality of plasma and of stored white cell (WBC)-reduced red cells (RBCs) from overnight-stored WB, cooled with either of the systems. STUDY DESIGN AND METHODS: Temperature curves following cooling with Compocool or CompoCool-WB were studied with a 25 percent glycerol solution as simulated WB. WB from voluntary donors was cooled with Compocool or CompoCool-WB, stored overnight at room temperature, centrifuged, and separated into components. WBC-reduced RBCs in SAGM were stored until Day 42 with measurement of in vitro parameters (n=23/group). RESULTS: Simulated WB reached a temperature of less than 25 degrees C after 2:15+/-1:04 hours for Compocool versus 1:39+/-0:38 hours for CompoCool-WB (p=0.02). On Day 35, RBCs had a hemolysis of 0.3+/-0.2 percent in both groups, and ATP levels were 3.3+/-0.5 and 3.6+/-0.5 micromol per g hemoglobin for Compocool and CompoCool-WB, respectively (not significant). Factor VIII content in plasma was 1.05+/-0.25 and 0.97+/-0.18 IU per mL for Compocool and CompoCool-WB, respectively. CONCLUSION: WB can be cooled to room temperature within 2 hours with both Compocool and CompoCool-WB butane-1,4-diol plates, improving temperature uniformity in WB donations. Application of either design for overnight storage of WB at room temperature had no adverse effects on the composition of subsequently prepared blood components.     

Automated blood component collection with the MCS 3p cell separator: evaluation of three protocols for buffy coat-poor and white cell- reduced packed red cells and plasma

BACKGROUND: Automated collection of blood components with a cell separator (MCS 3p, Haemonetics), was performed according to three protocols. STUDY DESIGN AND METHODS: The first protocol provided 2 units of fresh-frozen plasma (FFP); and one buffy coat-poor red cell (RBC) concentrate in additive solution. The second protocol included an additional in-line filtration of the RBC in a closed system after storage at 4 degrees C for 24 hours. In the third protocol, an additional platelet concentrate (PC) was recovered from the buffy coat. Cell counts and biochemical characterization of the RBCs (n=20 each) were determined on Days 0, 1, 14, 28, and 49. RESULTS: The RBC volume was 336 +/? 9 mL (first protocol), 337 +/? 7 mL (second protocol) and 293 +/? 12 mL (third protocol) with a hematocrit of 59 +/? 2, 53 +/? 3, and 61 +/? 5, percent respectively. On Day 49, hemolysis was 0.24 +/? 0.1 percent (first protocol), 0.33 +/? 0.32 percent (second protocol), and 0.38 +/? 0.1 percent (third protocol). The filtered RBC concentrate met the international standards for white cell-reduced RBCs. Filtration resulted in a clinically irrelevant increase of hemolysis. The in vitro RBC values (lactate dehydrogenase, 2-hydroxybutyrate dehydrogenase, hemolysis, potassium, 2,3 DPG, ATP) were at least equal to those in RBCs collected by conventional whole-blood donation. There is a trend toward extended preservation of 2,3 DPG in RBCs collected by apheresis. Two units of FFP could be collected with each donation (first protocol: 420 +/? 55 mL, 5.4 +/? 7 WBCs/microL, 6.5 +/? 5 × 10(3) platelets/microL; second protocol: 440 +/? 33 mL, 3 +/? 5.2 WBCs/microL, 32 +/? 12 × 10(3) platelets/microL; third protocol: 398 +/? 32 mL, 5 +/? 12 WBCs/microL; 3.4 +/? 3.5 × 10(3) platelets/microL). PCs prepared from the buffy coat collected by the third protocol contained 90 +/? 30 × 10(9) platelets in 88 +/? 14 mL of plasma. In vitro test results in these PCs were superior to those in PCs collected by conventional whole-blood donation. The procedure was well tolerated by all donors. No adverse reactions appeared. CONCLUSION: Erythroplasmapheresis with the MCS 3p cell separator is a useful alternative to conventional whole-blood donation and separation.     

Storage of interim platelet units for 18 to 24 hours before pooling: in vitro study

BACKGROUND: The Atreus 3C system (CaridianBCT) automatically produces three components from whole blood (WB), a red blood cell (RBC) unit, a plasma unit, and an interim platelet (PLT) unit (IPU) that can be pooled with other IPUs to form a PLT dose for transfusion. The Atreus 3C system also includes a PLT yield indicator (PYI), which is an advanced algorithm that provides an index that is shown to correlate well with the amount of PLTs that finally end up in the IPU bag. The aim of our in vitro study was to compare the effects of holding WB overnight versus processing WB fresh (2‐8 hr), both with 18‐ to 24‐hour storage of the IPUs before pooling into a transfusable PLT dose. STUDY DESIGN AND METHODS: WB was processed either fresh (within 8 hr after collection, Atreus F) or after overnight storage (14‐24 hr, Atreus S) without agitation at 22 ± 2°C. After a subsequent resting time of 18 to 24 hours on a flat‐bed shaker, five IPUs were selected for pooling with 200 mL of PAS II for in vitro quality assessments during a 7‐day storage period (n = 10 in each arm). IPUs were selected for pooling using the PYI of the Atreus 3C system. RESULTS: During storage, the glucose concentration was lower (p < 0.05) and the lactate concentration was higher (p < 0.05) in Atreus S pools, but no differences in the glucose consumption rate were noted. Adenosine triphosphate levels and hypotonic shock response reactivity were higher in Atreus S (p < 0.05). No significant differences in PLT counts, contents, mean PLT volume, lactate dehydrogenase, pO₂, pCO₂, extent of shape change, and CD62P between groups were detected. pH was maintained higher than 6.8 (Day 7). With exception of 2 units in the Atreus S arm, swirling remained at greater than 2 in all units at all times. CONCLUSION: Our results suggest that PLTs prepared from fresh or overnight‐stored WB and pooled after 18 to 24 hours meet necessary in vitro criteria without any relevant differences between both groups. Using the PYI, comparable yields can be obtained between WB processed within 2 to 8 hours and WB stored overnight.     

Coagulation parameters in apheresis and leukodepleted whole-blood plasma during storage

BACKGROUND: Thawing fresh-frozen plasma (FFP) may cause delay in delivery, and one approach to circumvent this is to store plasma at +4 degrees C. Thawed plasma is commonly discarded after a few days of storage, owing to the assumption that coagulation factor activity decreases to clinically unacceptable levels. STUDY DESIGN AND METHODS: Eighteen apheresis plasma (AP) units were collected from blood donors. The collected plasma was divided into two equal parts: one part frozen at -74 degrees C as FFP and one part stored at +4 degrees C as fresh liquid plasma (FLP). Thirty-nine units of whole blood (WB) were collected from blood donors and leukodepleted by inline filtration, followed by plasma separation. Twenty plasma units were frozen at -74 degrees C as FFP and 19 plasma units were stored at +4 degrees C as FLP for 28 days. Plasma aliquots were collected before freezing and immediately after thawing FFP and before and during storage of FLP at Days 14 and 28. Factor (F)V, FVIII, D-dimers, and C1-esterase inhibitor levels were assessed. RESULTS: No significant differences in coagulation factor levels were assessed between FLP prepared from AP and FLP prepared from WB. FV and FVIII levels decreased on average 25 and 50 percent, respectively, at Day 14 of storage. C1-esterase inhibitor and D-dimers levels were not affected. CONCLUSION: Leukodepleted apheresis and WB plasma stored for 14 days retain sufficient levels of FV and FVIII activity for maintenance of normal hemostasis and could therefore be considered useful in selected clinical situations.     

Phosphorylated Carbohydrate Intermediates of the Human Erythrocyte during Storage in Acid Citrate Dextrose:

Blood was stored at 4 C. in ACD without additive or supplemented with adenine, inosine, inosine-adenine, or adenosine. After six weeks of storage the carbohydrate intermediates of the erythrocytes were studied by column chromatography. Adenosine triphosphate (ATP) and 2,3-diphosphoglycerate (DPG) had disappeared from blood stored without additive or with adenine. Both ATP and DPG remained in erythrocytes stored with the other additives. Concentration of ATP was highest in blood stored with inosine-adenine, and the level of DPG was highest with the addition of adenosine to the storage solution.     

Toward a definition of “fresh” whole blood: an in vitro characterization of coagulation properties in refrigerated whole blood for transfusion

BACKGROUND: The hemostatic property of “fresh” whole blood (WB) has been observed in military application and cardiac surgery and is associated with reduced blood loss, transfusion requirements, and donor exposures. The time from donation to transfusion defining “fresh” has not been systematically studied. We undertook an in vitro study of coagulation properties of refrigerated WB stored for 31 days. STUDY DESIGN AND METHODS: Twenty‐one WB units were obtained from healthy volunteer donors and stored under standard AABB refrigerated conditions. Samples were obtained on the day after donation and again on Days 2, 4, 7, 11, 14, 17, 21, 24, and 31. Tests included complete blood count, pH, pO₂, pCO₂, glucose, lactate, thromboelastography (TEG), and platelet function by light transmission aggregometry (LTA). RESULTS: There was progressive decline in pH, pO₂, glucose, and sodium, but progressive increase in potassium, pCO₂, and lactate. TEG variables in all units were normal through Day 11; abnormal values in some variables in some units began on Day 14. Final aggregation levels exhibited no change from Day 1 to Day 21 with adenosine diphosphate and epinephrine, but a decline with collagen (Day 7) and ristocetin (Day 17). CONCLUSION: This in vitro study of coagulation properties demonstrates preservation of normal integrated coagulation function to a minimum of 11 days under standard conditions of refrigerated storage of WB for transfusion. These observations strongly suggest that the hemostatic quality of WB may extend beyond current transfusion practices. If confirmed clinically, this would increase availability and extend benefits of reduced donor exposure and transfusion requirements.     

Rejuvenation of irradiated AS-1 red cells

BACKGROUND: Gamma irradiation of blood components is used to prevent transfusion-associated graft-versus-host disease. The demand for irradiated blood components is increasing because of the increase in directed donation by family members. Irradiated units currently have a recommended maximum storage life of 28 days. Since in vivo recovery is related to red cell ATP levels, rejuvenation of stored irradiated units using a pyruvate-inosine phosphate-adenine additive was explored. STUDY DESIGN AND METHODS: Units of AS-1 red cells from 16 volunteer donors were divided into two equal volumes and one split unit from each was irradiated with 25 Gy. Ten units were irradiated on Day 5, 6, or 7 of 4 degrees C storage and 6 units were irradiated on Day 1 of 4 degrees C storage. All units were rejuvenated for 1 hour at 37 degrees C using a pyruvate-inosine-phosphate-adenine additive on Day 42 of 4 degrees C storage. Units were assayed for ATP, 2, 3 DPG and supernatant sodium, potassium, and glucose. RESULTS: ATP and 2, 3 DPG levels were restored equally well in irradiated and non-irradiated units. The previously reported irradiation-induced red cell potassium-sodium shift was demonstrated. Supernatant potassium and sodium levels did not reverse 1 hour after rejuvenation was completed. There was no significant difference in results between units irradiated on Day 1 or Day 5, 6, or 7. CONCLUSION: Red cell ATP and 2, 3 DPG levels were restored in irradiated AS-1 units stored at 4 degrees C for 42 days using a pyruvate-inosine-phosphate-adenine rejuvenation additive.     

Manufacture of red cells in additive solution from whole blood refrigerated for 5 days or remanufactured from red cells stored in plasma

Background and Objectives: To investigate methods for the production of red cell concentrates (RCC) in saline, adenine, glucose and mannitol (SAG‐M), from whole blood or red cells stored in plasma for 5 or 6 days and to provide evidence that exchange transfusion RCC in citrate phosphate dextrose (CPD) plasma or citrate, phosphate, dextrose, adenine (CPDA‐1) plasma are of comparable quality. Methods and Materials: Ten RCC in SAG‐M were produced following the remanufacture of red cells in CPD plasma on day 5/6 or after 5 days hold as leucodepleted CPD whole blood. In addition, 10 RCC in CPD plasma and 9 in CPDA‐1 plasma were stored without further processing. Units were assessed for red cell parameters including haemolysis, adenosine triphosphate (ATP), 2,3‐diphosphoglycerate (2,3‐DPG) and extracellular potassium. Results: Units in SAG‐M produced by remanufacture of RCC in plasma or by delayed manufacture of whole blood had comparable levels of haemolysis, ATP and 2,3‐DPG. Furthermore, these units underwent biochemical changes similar to reference SAG‐M units, with the exception of haemolysis which was greater at the end of shelf life and supernatant potassium which was lower following remanufacture. As expected, the decline in ATP was greater in red cells stored in CPD plasma compared with CPDA‐1 plasma. In general, units in CPD plasma were of similar quality at day 28 compared to those in CPDA‐1 plasma at day 35. Conclusions: RCC produced following the remanufacture of RCC in plasma or the delayed manufacture of whole blood are of acceptable in vitro quality and should be assigned the same shelf life as standard RCC in SAG‐M.     

Analysis of prolonged storage on coagulation Factor (F)V,FVII, and FVIII in thawed plasma: is it time to extend the expiration date beyond 5 days?

BACKGROUND: According to AABB standards, fresh‐frozen plasma (FFP) should be thawed at 30 to 37°C and expire after 24 hours. An increase in the aggressive management of trauma patients with thawed plasma has heightened the risk of plasma waste. One way to reduce plasma waste is to extend its shelf life, given that the full range of therapeutic efficacy is maintained. We evaluated the effect of prolonged storage at 1 to 6°C on the activity of Factor (F)V, FVII, and FVIII in plasma thawed at 37 or 45°C. STUDY DESIGN AND METHODS: Group O plasma from healthy donors (n = 20) was divided into 10 pairs and frozen and stored at not more than ?18°C. One sample from each pair was thawed at 37 or 45°C, and all were stored at 1 to 6°C. Samples were analyzed for FV, FVII, and FVIII activity on Days 0, 5, 10, 15, and 20. RESULTS: Plasma thawing time was 17% less at 45°C than at 37°C. No differences were observed between thawing groups in coagulation activity of FV, FVII, and FVIII during the 20‐day storage period (p > 0.12). In both groups, the activity of FV and FVIII decreased over time but remained within a normal range at 10 days. CONCLUSION: Although levels of plasma clotting factors are reduced in storage, therapeutic levels of FV and FVIII are maintained in thawed plasma stored for up to 10 days at 1 to 6°C. Thawing of FFP at 45°C decreases thawing time but does not affect the activity of FV, FVII, and FVIII.     

Storage of aliquots of apheresis platelets for neonatal use in syringes with and without agitation

BACKGROUND: To facilitate volume control in neonates, platelets (PLTs) are aliquoted and stored for short periods in non–gas‐permeable syringes before infusion. Although agitation of PLTs during storage in gas‐permeable bags is performed to maintain their quality, the effect of syringe agitation during storage is unknown. STUDY DESIGN AND METHODS: Double apheresis PLTs (n = 6) were collected and split, providing two identical products. On Days 2 and 4 of storage, aliquots from one bag of each pair were transferred to two syringes and stored for 6 hours on flatbed agitator or were left at 20 to 24°C without agitation. A series of in vitro tests was performed on Days 0, 2 (Hours 0 and 6), and 4 (Hours 0 and 6). Control samples were obtained from the second matched bag that was stored on the agitator. Data were analyzed by one‐way analysis of variance with differences considered significant if the p value was less than 0.05. RESULTS: Comparable results for several PLT variables were obtained with or without agitation of the syringes. On Day 4 Hour 6, pH values were 7.18 ± 0.12 (agitated syringes) and 7.19 ± 0.1 (nonagitated syringes), and extent of shape change and hypotonic shock response measurements were not significantly different between agitated syringes and nonagitated syringes (23.7 ± 6.4 and 74.3 ± 9.8% vs. 23.3 ± 5.4 and 76.0 ± 7.6%), respectively. CONCLUSION: Based on in vitro testing, apheresis PLT aliquots can be stored in syringes for at least 6 hours without agitation before transfusions.