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.     

The influence of maintaining the correct whole blood-to-anticoagulant ratio during donation on the quality of leukoreduced whole blood

BACKGROUND: It is unclear whether maintaining the correct whole blood‐to‐anticoagulant (WB : AC) ratio during collection can improve the quality of red blood cell (RBC)‐containing blood products to a clinically relevant degree. STUDY DESIGN AND METHODS: A total of 2 × 20 CPDA‐1 leukoreduced whole blood (WB) units suspended in CPDA‐1 were investigated. In one group, AC was continuously added to the donated blood, maintaining the correct WB : AC ratio during collection, using a new drawing device (MacoPharma ABC). In the other group, WB units were produced conventionally. Adenosine triphosphate (ATP), 2,3‐diphosphoglycerate, free hemoglobin (Hb), potassium, glucose, lactate, pH, and variables of coagulation were determined on Days 1, 7, 21, 35, 42, and 49 of storage. Variables of RBC deformability and aggregability were determined using a laser‐assisted optical rotational cell analyzer. RESULTS: The ABC and conventional group showed comparable unit volumes of 525 (SD, 5.3) mL versus 524 (SD, 10.2) mL and Hb content of 65.9 (SD, 5.1) g/unit versus 67.5 (SD, 7.8) g/unit, but higher variation after conventional blood drawing (p = 0.006 and p = 0.07, respectively) was observed. During storage, none of the measured quality variables were significantly different between the groups. Mean (SD) ATP was 2.33 (0.41) µmol/g Hb versus 2.24 (0.39) µmol/g Hb after 42‐day storage. Deformability was not different (p = 0.44), whereas the extent of the aggregability was higher in the conventional group (p = 0.04). CONCLUSION: The ABC device provided a better standardized blood product but did not improve RBC storage variables or plasma quality. It slightly reduced RBC aggregability during storage. Excess AC at the beginning of a donation appears not to significantly affect RBC storage in conventional blood drawing.     

In vitro and in vivo evaluation of a whole blood platelet‐sparing leukoreduction filtration system

BACKGROUND: In‐line leukoreduction (LR) filters decrease adverse clinical sequelae caused by residual white blood cells (WBCs). Such filtration, however, can remove platelets (PLTs) needed for production of PLT concentrates (PCs). This study measured in vitro and in vivo efficacy of a new whole blood PLT‐sparing LR filter (WBPSF) system that performs whole blood (WB) LR using a single closed‐system filtration step. The WBPSF provides three final LR products: AS‐5 red blood cells (RBCs), citrate‐phosphate‐dextrose (CPD) PLTs, and CPD plasma. STUDY DESIGN AND METHODS: Volunteers (n = 59) donated WB processed using the WBPSF system. WB filtration time was recorded, and LR WB was processed into AS‐5 LR RBCs, CPD LR PLTs, and LR plasma. Final components were assayed for in vitro indices, and in vivo characteristics for LR AS‐5 RBCs and CPD PLTs were assayed after radiolabeling. RESULTS: WB filtration time averaged 37 minutes. Transfusion products obtained after WBPSF met all in vitro and in vivo Food and Drug Administration (FDA) requirements. Radiolabeling of LR AS‐5 RBCs after WBPSF showed a 24‐hour RBC recovery of 81.3 ± 5.3% after 42 days of storage. In vivo dual ¹¹¹In/⁵¹Cr radiolabeling of PCs manufactured using WBPSF showed a Day 5 recovery ratio of 80 ± 19% versus fresh autologous PLTs and a survival ratio of 81 ± 17% that of fresh autologous PLTs. CONCLUSION: All WBPSF‐derived transfusion products met or exceeded in vitro and in vivo FDA guidelines. This filtration system is suitable for routine blood center or hospital use in the production of LR AS‐5 RBCs, CPD PLTs, and CPD plasma.     

白膜法制备浓缩血小板后悬浮红细胞质量探讨

目的 观察白膜法制备浓缩血小板后的悬浮红细胞质量。方法 对白膜法制备浓缩血小板后的悬浮红细胞质量与常规方法制备的进行比较。检测两者血容量、血红蛋白（Hb）、白细胞（WBC）、血小板（PLT），并比较相同Hb含量时两者的WBC、PLT含量。结果 两者血容量、Hb、WBC、PLT含量及相同Hb量时WBC、PLT含量的差异均有统计学意义（P〈0．01）。结论 白膜法制备浓缩血小板后的悬浮红细胞的WBC、PLT含量较少，输血更安全；但血容量、Hb含量明显低于常规方法制备的悬浮红细胞，血液有效成分含量减少。     

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.     

Platelet-white blood cell (WBC) interaction, WBC apoptosis, and procoagulant activity in stored red blood cells

BACKGROUND: Nonleukoreduced units of red blood cells (RBCs) contain activated platelets (PLTs) that interact with white blood cells (WBCs) and may promote inflammation and thrombosis in the recipient. The aim of this study was to characterize PLT‐WBC interactions (PLT‐WBC aggregates [PLAs]), WBC apoptosis, WBC death, and the development of procoagulant activity in RBCs during storage. STUDY DESIGN AND METHODS: RBCs were prepared from volunteer donor blood and stored. Samples were analyzed with flow cytometry between Days 1 and 15 to measure PLT‐monocyte aggregate (PMA) and PLT‐neutrophil aggregate (PNA) formation, WBC apoptosis (annexin V binding), and cell death (binding of 7‐aminoactinomycin D). Procoagulant activity in the supernatant of four RBC preparations was assessed between Days 1 and 39 using a clotting assay with and without the addition of an inhibitory anti‐tissue factor (TF) antibody, αTF‐5. RESULTS: PLA formation was extensive and maximal on Day 3 of storage (PNA, 23 ± 13%; PMA, 93 ± 4%; n = 6). Apoptosis was progressive throughout storage, with 95 ± 4% of neutrophils and 73 ± 19% of monocytes binding annexin V on Day 15. Cell death became measurable after apoptosis. Procoagulant activity was observed in all RBCs but with varying temporal patterns. It was partially TF dependent and removed with high‐speed centrifugation, suggestive of an association with microparticles. CONCLUSION: The activation of PLTs during the storage of RBCs induces PLA formation that precedes WBC apoptosis and death. Procoagulant activity, likely associated with microparticles derived from apoptotic WBCs, may contribute to adverse effects of stored, nonleukoreduced RBCs.     

Effect on the quality of blood components after simulated blood transfusions using volumetric infusion pumps

BACKGROUND: It is unknown whether the use of volumetric infusion pumps for the transfusion of red blood cells (RBCs) or platelet (PLT) concentrates (PCs) affects the quality of the blood components. We therefore investigated the in vitro quality of these components after use of infusion pumps. STUDY DESIGN AND METHODS: Ten different volumetric infusion pumps were used to simulate transfusion with RBCs and PCs. To prevent donor‐dependent differences multiple units were pooled and divided into equal portions. The storage time of RBCs was 30 to 35 days (n = 10 experiments), and for PCs, either 2 (n = 5) or 7 days (n = 5). For RBCs an infusion rate of 100 or 300 mL/hr was used, and for PCs, 600 mL/hr. Transfusions without an infusion pump served as a reference. RESULTS: None of the infusion pumps induced an increase of free hemoglobin, annexin A5 binding, or formation of echinocytes in RBCs compared to reference units. In 2‐ and 7‐day‐old PCs no effect was shown on PLT concentration, annexin A5 binding, mean PLT volume, and morphology score compared to the reference. The CD62P expression of 2‐day‐old PCs was significantly lower after transfusion compared to the reference, that is, 11.7 ± 2.1% versus 8.1 ± 1.3% (p < 0.01). CONCLUSION: There was no adverse effect on the in vitro quality of RBCs or PCs after simulated transfusion using volumetric infusion pumps. A decrease in PLT activation was observed, which can probably be explained by capturing of activated or damaged PLTs in the 200‐µm filter present in the infusion system.     

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.     

Switching iron‐deficient whole blood donors to plateletpheresis

BACKGROUND: Iron deficiency is a frequent side effect of whole blood (WB) donation. In contrast, less red blood cell loss and therefore less iron loss results from plateletpheresis. STUDY DESIGN AND METHODS: WB donors presenting a decrease in either hemoglobin (Hb) or ferritin levels were offered to switch to plateletpheresis with or without iron supplementation. We analyzed the effect of this intervention on deferral rates for an insufficient Hb level in 168 donors. Further, we assessed how this intervention affected Hb and ferritin levels, anemia occurrence, and platelet (PLT) concentrate yields in the donors who presented at least four successive times for thrombapheresis. RESULTS: Switching WB donors to repetitive plateletpheresis procedures resulted in an increase of median Hb (+12 g/L, p < 0.001) and ferritin (+15.5 ng/mL, p = 0.002) values. Anemia and deferral rates were reduced by 23% (p = 0.004) and 13% (p < 0.001). Between high‐ and low‐frequency apheresis donors, no significant differences in Hb and ferritin levels were found. Similarly, discrepancies in Hb and ferritin values between donors that adopted iron supplementation and those who did not were insignificant. The median PLT concentrate yield was 5.43 × 10¹¹ PLTs. CONCLUSION: Switching iron‐deficient WB donors to plateletpheresis was an effective intervention that permitted us to correct low Hb and ferritin levels while retaining donors in our pool.     

Hemolysis and red blood cell mechanical fragility in shed blood after total knee arthroplasty

BACKGROUND: There are several options for the salvage of postoperative shed red blood cells (RBCs). This study compared the characteristics of the returned RBCs collected using two different devices: one that washes and one that does not wash the collected RBCs. STUDY DESIGN AND METHODS: Forty patients undergoing first‐time total knee arthroplasty consented to participate. Twenty patients were operated on by a surgeon who routinely uses a device that does not wash the shed RBCs (unwashed group), the other 20 patients were operated on by surgeons who routinely use a device that washes and concentrates the collected RBCs (washed group). A small quantity of postprocessing RBCs were collected immediately before reinfusion and the amount of plasma‐free hemoglobin (PFHb), and the mechanical fragility index (MFI) of the returned RBCs were determined. RESULTS: The patients in both groups were well matched for age, sex, and length of stay. The mean percent hemolysis of the returned RBCs was not different between the unwashed and washed groups (1.22 ± 0.30 vs. 1.24 ± 0.42, p = 0.895), while the mean total amount of returned PFHb was not different (0.51 ± 0.12 g vs. 0.55 ± 0.35 g, p < 0.615). The ratio of total PFHb:total returned Hb was significantly lower for the washed group (0.0087 ± 0.0023 vs. 0.0035 ± 0.0011, p < 0.0001). The MFI was higher in the washed group (1.71 ± 0.55 vs. 0.53 ± 0.42, p < 0.001). CONCLUSIONS: The washing device returned more Hb to the patients relative to the amount of free Hb.     

Comparison of two sterile connection devices and the effect of sterile connections on blood component quality

BACKGROUND: Sterile connection devices (SCDs) are used to connect pieces of polyvinylchloride tubing between blood bag systems. After observing a slight decrease in inner diameter of tubing welded with the CompoDock S2 SCD, the effect of welded tubing on storage characteristics of white blood cell (WBC)-reduced red blood cells (RBCs) and platelet (PLT) concentrates was studied. Welds were made with Terumo SCD (T-SCD) or CompoDock S2, and unwelded tubing served as reference. STUDY DESIGN AND METHODS: Three WBC-reduced RBC units or 3 PLT concentrates were pooled and divided to prevent donor-dependent differences. The units were transferred 10 times over (1) tubing with a T-SCD weld, (2) a CompoDock S2 weld, or (3) unwelded tubing. RBCs were stored for 42 days and free hemoglobin (Hb) was measured; PLT concentrates were stored for 8 days and CD62P expression was measured, as markers for blood component quality (n = 10 paired experiments). RESULTS: WBC-reduced RBC units had similar hemolysis at the end of storage: 0.47 +/- 0.28, 0.47 +/- 0.35, and 0.49 +/- 0.38 percent of total Hb, for tubing with a T-SCD weld, a CompoDock S2 weld, or no weld, respectively (not significant). CD62P expression of stored WBC-reduced PLT concentrates was not significantly different between the groups: 20.3 +/- 5.1, 19.8 +/- 5.1, and 22.3 +/- 9.8 percent for tubing with a T-SCD weld, a CompoDock S2 weld, or no weld, respectively. CONCLUSION: The quality of blood components, measured as RBC hemolysis and platelet CD62P expression, is not adversely affected by the presence of a sterile connection in the tubing, made by either the CompoDock S2 or the T-SCD.     

In vivo viability of stored red blood cells derived from riboflavin plus ultraviolet light-treated whole blood

BACKGROUND: A novel system using ultraviolet (UV) light and riboflavin (Mirasol System, CaridianBCT Biotechnologies) to fragment nucleic acids has been developed to treat whole blood (WB), aiming at the reduction of potential pathogen load and white blood cell inactivation. We evaluated stored red blood cell (RBC) metabolic status and viability, in vitro and in vivo, of riboflavin/UV light–treated WB (IMPROVE study). STUDY DESIGN AND METHODS: The study compared recovery and survival of RBCs obtained from nonleukoreduced WB treated using three different UV light energies (22, 33, or 44 J/mL_RBC). After treatment, WB from 12 subjects was separated into components and tested at the beginning and end of component storage. After 42 days of storage, an aliquot of RBCs was radiolabeled and autologously reinfused into subjects for analysis of 24‐hour recovery and survival of RBCs. RESULTS: Eleven subjects completed the in vivo study. No device‐related adverse events were observed. By Day 42 of storage, a significant change in the concentrations of sodium and potassium was observed. Five subjects had a 24‐hour RBC recovery of 75% or more with no significant differences among the energy groups. RBC t_1/2 was 24 ± 9 days for the combined three groups. Significant correlations between 24‐hour RBC recovery and survival, hemolysis, adenosine triphosphate (ATP), and CO₂ levels were observed. CONCLUSIONS: This study shows that key RBC quality variables, hemolysis, and ATP concentration may be predictive of their 24‐hour recovery and t_1/2 survival. These variables will now be used to assess modifications to the system including storage duration, storage temperature, and appropriate energy dose for treatment.     

二甲基亚砜在人红细胞冻干前负载海藻糖过程中的作用

目的研究人红细胞冻干保存前负载海藻糖过程中二甲基亚砜(DMSO)的作用,优化红细胞负载缓冲液配方。方法实验组以浓缩红细胞25份(10ml/份)负载海藻糖,负载缓冲液中添加DMSO;对照组25份负载海藻糖,负载缓冲液中未添加DMSO。37℃条件下孵育8h后,分别检测两组红细胞胞内海藻糖负载量、胞外游离血红蛋白水平、ATP含量、红细胞变形性,并利用流式细胞术检测负载后红细胞膜的完整性。结果实验组与对照组红细胞的胞内海藻糖负载量分别为(57.033±4.883)mmol/L,(49.184±4.858)mmol/L(P<0.05);胞外游离血红蛋白浓度分别为(4.131±0.473)g/L,(5.410±0.501)g/L(P<0.05);ATP浓度分别为(3.874±0.426)μmol/g Hb,(3.358±0.306)μmol/g Hb(P<0.05);红细胞变形指数分别为0.330±0.0211,0.277±0.0232(P<0.01);红细胞胞膜PS表达率分别为(5.04±0.495)%,(8.69±0.862)%(P<0.01)。结论DMSO在红细胞负载海藻糖过程中可有效增加胞内海藻糖负载量,并显著改善负载缓冲液对红细胞胞膜的高渗损伤,更好地发挥海藻糖对红细胞的保护作用。     

Multicenter evaluation of a whole-blood filter that saves platelets

BACKGROUND: Whole-blood (WB) leukoreduction filters in current use retain the majority of PLTs. A new whole-blood filter, which retains significantly fewer of the PLTs (or saves PLTs [WB-SP]), has been developed. The performance characteristics of the WB-SP filter have been evaluated in a multicenter study. STUDY DESIGN AND METHODS: A total of 617 units of WB was collected into quadruple bag sets with an integrated WB-SP filter, leukoreduced, and processed into leukoreduced RBCs (LR-RBC), plasma (LR-PL), and buffy coats (LR-BC) from which, pooled, leukoreduced, PLT concentrates (LR-PCs) were produced. Recovery, yield, and residual WBCs were assessed in prepared blood components. RESULTS: The median residual WBC number in the LR-RBCs was 0.05 x 10(6) (range, <0.05-3.8), exceeding 1 x 10(6) in 0.6 percent of the units. Median Hb content in LR-RBC was 50 g (range, 34-72), reflecting a final RBC recovery of 81 +/- 6 percent. The median WBC content of the LR-PC was 0.05 x 10(6) (range, <0.05-0.28), with none exceeding 1 x 10(6). The median PLT content of the LR-PC, per individual donation, was 6.4 x 10(10) (range, 4.1-10.7), representing a final recovery of 62 +/- 10 percent. The mean FVIII activity was 104 +/- 25 percent and 83 +/- 11 percent in plasma separated from fresh or overnight stored WB, respectively. CONCLUSION: Use of the WB-SP filter makes it possible to obtain three leukoreduced blood components with only one filtration step. The WB-SP filter showed good leukoreduction performance and recovery of all blood components including PLTs.     

Level of platelet-derived cytokines in leukoreduced red blood cells is influenced by the processing method and type of leukoreduction filter

BACKGROUND: In contrast to the well-documented effect of white blood cells on the quality of red blood cells (RBCs), the effect of platelets (PLTs) has received little consideration. In this study, the PLT content and level of PLT-derived cytokines in RBCs prepared using different types of leukoreduction methods were investigated.
STUDY DESIGN AND METHODS: Buffy coat–poor RBCs and five types of leukofiltered (LF) RBCs, including RBCs prepared with a whole blood (WB) PLT-saving filter, were prepared and stored according to standard blood bank conditions. PLT content was measured on Day 1, and levels of PLT-derived cytokines were measured by enzyme-linked immunosorbent assay at nominated timepoints during 42 days of storage.
RESULTS: The PLT content of leukoreduced RBCs varied widely depending on the processing method and/or leukoreduction filter used, with some types of RBCs containing very low PLT counts while other units contained PLT counts comparable to those of unprocessed WB. The PLT content of RBCs directly influenced the concentration and accumulation of PLT-derived cytokines. Several PLT-derived factors exhibited significant accumulation throughout 42 days of storage. RBCs with high PLT content exhibited concentrations of RANTES (CCL5) and soluble CD40 ligand equivalent to those previously reported to show significant biologic and clinical effects.
CONCLUSION: The PLT content and levels of PLT-derived cytokines in leukoreduced RBCs are influenced by the processing method and types of leukoreduction filters used. It may be inappropriate to consider LF-RBCs prepared with different types of leukoreduction filters as equivalent products based on their differing levels of PLT factors.     

Gravity sedimentation of granulocytapheresis concentrates with hydroxyethyl starch efficiently removes red blood cells and retains neutrophils

BACKGROUND: Transfusion of granulocytapheresis concentrates can be limited by the volume of incompatible donor red blood cells (RBCs) in the component. Efficient reduction of RBCs in granulocyte units would result in safe transfusion of RBC‐incompatible units. STUDY DESIGN AND METHODS: Granulocyte concentrates were collected by continuous‐flow apheresis from granulocyte–colony‐stimulating factor (G‐CSF) and dexamethasone‐stimulated volunteer donors, with 6% hydroxyethyl starch (HES) added continuously during apheresis as a RBC sedimenting agent to enhance granulocyte collection efficiency. After collection, the component was placed in a plasma extractor for 4 hours. A sharp line of demarcation between the starch‐sedimented RBCs and the granulocyte‐rich supernatant developed, and the supernatant was transferred to a sterilely docked transfer pack. RBC reduction and white blood cell recovery were determined. RESULTS: Gravity sedimentation was performed on 165 granulocyte concentrates. Mean sedimentation time was 267 minutes (range, 150‐440 min). RBC depletion was 92% (range, 71%‐99%) with mean residual RBC content of 3.2 ± 1.4 mL. Twelve percent of components contained less than 2 mL of RBCs. Mean granulocyte and platelet (PLT) recoveries were 80 and 81%, respectively. There were no transfusion reactions or signs of hemolysis after transfusion of 66 RBC‐incompatible granulocyte concentrates (RBC volume, 1.6‐8.2 mL). The remaining concentrates were used for topical or intrapleural applications. CONCLUSIONS: RBCs were significantly reduced and granulocytes and PLTs effectively retained in G‐CSF/steroid–mobilized granulocyte components collected with HES and processed by gravity sedimentation. This procedure allows safe transfusion of RBC‐incompatible sedimented granulocyte units and may be used to expand the pool of available granulocyte donors for specific recipients.     

Platelet concentrates prepared after a 20- to 24-hour hold of the whole blood at 22°C

BACKGROUND: The Food and Drug Administration (FDA) requires that red blood cells must be refrigerated within 8 hours of whole blood collection. Longer storage of whole blood at 22°C before component preparation would have many advantages. STUDY DESIGN AND METHODS: Two methods of holding whole blood for 20 to 24 hours at room temperature were evaluated, refrigerated plates or a 23°C incubator. After extended whole blood storage, platelet (PLT) concentrates were prepared from PLT‐rich plasma on Day 1 postdonation, and the PLTs were stored for 6 more days. On Day 7 of PLT storage, blood was drawn from each subject to prepare fresh PLTs. The stored and fresh PLTs were radiolabeled and transfused into their donor. RESULTS: Eleven subjects' whole blood was stored using refrigerated butanediol plates (Compocool, Fresenius), and 10 using an incubator. Poststorage PLT recoveries averaged 47 ± 13% versus 53 ± 11% and survivals averaged 4.6 ± 1.7 days versus 4.7 ± 0.9 days for Compocool versus incubator storage, respectively (p = NS). With all results, poststorage PLT recoveries averaged 75 ± 10% of fresh and survivals 57 ± 13% of fresh; PLT recoveries met FDA guidelines for poststorage PLT viability but not survivals. CONCLUSION: Seven‐day poststorage PLT viability is comparable when whole blood is stored for 22 ± 2 hours at 22°C using either refrigerated plates or an incubator to maintain temperature before preparing PLT concentrates.     

In vitro characteristics of red blood cell concentrates prepared from under- and overcollected units of whole blood and from a paediatric blood bag system

The problem of how to deal with red blood cell concentrates (RBCs) prepared from under- or overcollected units of whole blood (WB) and how to collect blood from underweight persons arises in the context of autologous predeposit. To determine the quality of RBCs stored in PAGGS-M additive solution prepared from under- and overcollected units of whole blood and of PAGGS-M RBCs prepared from a paediatric 250-mL top outlet blood bag system we measured blood picture, haemolysis, K+, pH, ATP and 2,3-DPG on days 0, 10, 20, 30, 40 and 49 of storage. The volume of WB collected ranged from 150 to 600 mL in 50-mL increments (4 units per volume). Haemolysis was under 0.8% on day 49 in all RBCs prepared from WB donations between 200 mL and 600 mL. However, the day 49 haemolysis level of standard RBCs prepared from 450 mL of WB (0.15 +/- 0.03%) was reached earlier in RBCs from under- and overcollected units of whole blood. 2,3-DPG levels decreased rapidly between days 10 and 20 in all RBCs studied. RBCs from 450-mL donations showed acceptable ATP maintenance after 49 days (70.4% of day 0 value), while all other RBC ATP levels were below 50% of the day 0 level on day 49. In vitro quality data of RBCs prepared from a 250-mL donation in the paediatric blood bag system after storage for about 25 days were comparable to those after 49 days of storage of standard RBCs. Our results suggest that it is feasible to transfuse PAGGS-M RBCs prepared from under- as well as overcollected units of WB in the autologous setting. However, we strongly recommend shortening the storage period of such RBCs to maintain the quality level of standard RBCs.     

The effect of storing whole blood at 22 degrees C for up to 24 hours with and without rapid cooling on the quality of red cell concentrates and fresh-frozen plasma

BACKGROUND: Storage of whole blood (WB) for less than 24 hours at ambient temperature is permitted in Europe, but data directly comparing storage with and without active cooling are lacking, which was investigated and compared to current standard methods. STUDY DESIGN AND METHODS: WB was stored in one of four different ways for 24 hours after donation before processing on Day 1 to red cell concentrates (RCCs) in saline‐adenine‐glucose‐mannitol and fresh‐frozen plasma (FFP; n = 20 each): 1) at 22°C in plastic trays, 2) in cooling devices (Compocool II, NPBI), 3) at 4°C, or 4) processed from WB without storage less than 8 hours from donation (Day 0). RESULTS: 2,3‐Diphosphoglycerate (2,3‐DPG) in RCCs were lower after ambient storage compared with those processed on Day 0 or after 4°C storage. Rapid cooling slowed the loss of 2,3‐DPG but levels were undetectable by Day 21 with any method. On Day 42 of RCC storage, there was no significant difference between storage methods in levels of adenosine triphosphate or hemolysis. Potassium levels were lower in RCCs from WB stored at ambient compared with those produced on Day 0, regardless of the use of cooling plates. FFP produced from WB on Day 0 or after storage at ambient with or without active cooling met UK specifications (>75% of units >0.70 IU/mL Factor VIII). CONCLUSION: These data suggest that RCCs and FFP produced from WB that has been stored at ambient temperature with or without active cooling are of acceptable quality compared with those produced using current standard methods in the United Kingdom.     

Characterization of blood components prepared from whole-blood donations after a 24-hour hold with the platelet-rich plasma method

BACKGROUND: The preparation of platelet (PLT) concentrates (PCs) from PLT-rich plasma (PRP) requires that whole blood (WB) be processed within 8 hours of collection. Increasing WB storage time to 24 hours would be logistically attractive. This study compares the in vitro quality of blood components prepared from WB stored for 8 and 24 hours at room temperature before processing with the PRP method. STUDY DESIGN AND METHODS: WB units were collected from ABO-matched blood donors. To reduce individual variations, paired donations were drawn in parallel, pooled, and split back in the collection bag. One unit was held for 6 to 8 hours and the other for 22 to 24 hours at 20 to 24 degrees C. Prestorage leukoreduced components were prepared with the PRP as intermediate product and analyzed during storage. RESULTS: RBC units prepared after an 8- or 24-hour hold were comparable in terms of hemolysis, sodium, pH, and ATP levels. RBC 2,3- diphosphoglycerate (2,3-DPG) was significantly lower in RBCs prepared from 24-hour hold donations immediately after processing but not after 20 days of storage. Residual white blood cells were approximately fivefold higher (p < 0.05) in 24-hour RBC units. For PCs, measurements for glucose, ATP, lactate, pH, extent of shape change, hypotonic shock response, and CD62p activation were similar. No differences were observed in the von Willebrand factor, factor (F)V, FVIII, and fibrinogen content of fresh-frozen plasma. CONCLUSIONS: The decrease in FVIII and RBC 2,3-DPG can be acceptable as a compromise to improve blood component logistics, but leukoreduction efficiency must be improved before considering the adoption of an overnight storage of WB before PRP processing.