Method for storing whole blood and composition thereof

Abstract

The present application relates to methods and compositions for storing whole blood, methods and compositions for improving clinical outcomes in trauma patients receiving blood transfusions, and methods and compositions for improving clinical outcomes in cancer patients receiving blood transfusions.

Description

[0001] This application is a divisional application of the Chinese patent application with application date of May 18, 2016, application number "201680036477.2", and invention name "Method for storing whole blood and its composition". The original application is the Chinese national phase application of international application PCT / US2016 / 033151. Technical Field

[0002] The present disclosure relates to methods for improving the quality of whole blood that can be transfused into patients. Anaerobic storage of whole blood provides reduced cytokine levels and increased levels of 2,3-diphosphoglycerate (2,3-DPG) and adenosine triphosphate (ATP). The improved blood composition can be used for transfusion into cancer and trauma patients. Background Art

[0003] When stored in a conventional manner, stored blood undergoes a steady deterioration associated with various storage insults, including hemolysis, hemoglobin degradation, and reduced ATP and 2,3-DPG concentrations. When transfused into a patient, the effects of this steady deterioration during storage manifest as a reduction in 24-hour in vivo recovery, for example. Due to these and other medical sequelae of stored blood, various methods have been developed to minimize the effects of storage on blood and improve medical outcomes. See, for example, Zimring et al., “Established and theoretical factors to consider in assessing the red cell storage lesion” in Blood, 125: 2185-90 (2015).

[0004] A variety of approaches have been developed to minimize storage damage and improve transfusion outcomes. One approach is to develop additive solutions during storage. Examples of this approach include: U.S. Patent No. 4,769,318 to Hamasaki et al. and U.S. Patent No. 4,880,786 to Sasakawa et al., which are directed to additive solutions for blood preservation and activation. For example, (obtained from Citra Lab LLC, Braintree, MA) is added to blood immediately prior to transfusion or prior to freezing (i.e., at -80°C with glycerol) after refrigeration (i.e., at 4°C) to extend storage time. U.S. Patent No. 6,447,987 to Hess et al. relates to an additive solution for cold storage of human red blood cells. Another approach is to freeze the blood and prevent the development of storage damage. Storage of frozen blood is known in the art, but such frozen blood has limitations. U.S. Patent No. 6,413,713 to Serebrennikov relates to a method for storing blood at temperatures below 0°C. See Chaplin et al., "Blood Cells for Transfusion," Blood, 59: 1118-20 (1982), and Valeri et al., "The survival, function, and hemolysis of human RBCs stored at 4 degrees C in additive solution (AS-1, AS-3, or AS-5) for 42 days and then biochemically modified, frozen, thawed, washed, and stored at 4 degrees C in sodium chloride and glucose solution for 24 hours," Transfusion, 40: 1341-5 (2000). Another approach involves the container for blood storage provided by Sato et al., U.S. Patent No. 4,837,047.

[0005] One approach that has proven successful in improving the quality of blood and expanding its utility is through the depletion of oxygen and storage under anaerobic conditions. U.S. Patent No. 5,624,794 to Bitensky et al., U.S. Patent No. 6,162,396 to Bitensky et al., and U.S. Patent No. 5,476,764 to Bitensky are directed to the storage of red blood cells under hypoxic conditions. U.S. Patent No. 5,789,151 to Bitensky et al. is directed to a blood storage additive solution. Benefits of storing blood under hypoxic conditions include increased ATP and 2,3-DPG levels and reduced hemolysis. Storing blood under hypoxic conditions can also result in reduced levels of microparticles, reduced deformability losses, reduced lipid and protein oxidation, and higher post-transfusion survival rates compared to blood stored under conventional conditions.

[0006] US Patent No. 6,162,396 to Bitensky et al. (the '396 patent) discloses an anaerobic storage bag for blood storage comprising an oxygen-impermeable outer layer, an oxygen-permeable red blood cell (RBC)-compatible inner layer with an oxygen scrubber disposed between the inner and outer layers.

[0007] While the effects of oxygen depletion on packed red blood cells have been explored, the effects of hypoxia on whole blood have not been reported. This lack of research on whole blood deoxygenation may be due in part to the anticipated deleterious effects of oxygen deprivation on platelets. More specifically, given the crucial role of platelets in the coagulation process, there is concern that decreased platelet function could lead to coagulopathy and adverse clinical outcomes.

[0008] The storage of platelets has been extensively studied to determine the most favorable conditions, including temperature, pH, O2, and CO2 concentrations. The result of this work is that stored platelets persist in the recipient after transfusion, and platelets require oxygen and room temperature storage. Murphy and Gardner noted in 1975 that unwanted morphological changes are associated with reduced oxygen consumption. See Murphy et al., “Platelet storage at 22 degrees C: role of gas transport across plastic containers in maintenance of viability,” Blood 46(2): 209-218 (1975). The authors observed that increased oxygen uptake allows aerobic metabolism (oxidative phosphorylation), resulting in a decrease in lactate production. Increased lactate production at low PO2 levels is consistent with the Pasteur effect. Moroff et al. noted that continued oxygen consumption is required to maintain the pH of stored platelets at pH 7. See Moroff et al., "Factors Influencing Changes in pH during Storage of Platelet Concentrates at 20-24°C," Vox Sanguinis 42(1):33-45 (1982). Specially customized container systems are permeable to carbon dioxide and oxygen to prevent a fatal drop in pH. As shown in Kakaiya et al., "Platelet preservation in large containers," Vox Sanguinis 46(2):111-118 (1984), the reason for maintaining platelet quality is due to improved gas exchange conditions and increased surface area available for gas exchange. The importance of maintaining oxygen levels during platelet storage has led to the development of gas permeable containers and the storage of platelets in oxygen-enriched atmospheres. See U.S. Patent No. 4,455,299, issued to Grode on June 19, 1984. The importance of oxygen to the viability of stored platelets is reinforced because the content of lactate increases 5-8 times in an oxygen-deficient environment. See Kilkson et al., "Plateletmetabolism during storage of platelet concentrates at 22 degrees C." Blood 64(2): 406-14 (1984).Wallvik et al., "Platelet Concentrates Stored at 22°C Need Oxygen The Significance of Plastics in Platelet Preservation," Vox Sanguinis 45(4):303-311 (1983), disclose that maintaining oxygen for the first five days of storage is critical for platelet preservation. Wallvik and colleagues also showed that the maximum number of platelets that can be successfully stored for five days can be predicted based on the oxygen diffusion capacity of the storage bag. See Wallvik et al., "The platelet storage capability of different plastic containers," Vox Sanguinis 58(1):40-4 (1990). By providing a blood bag with adequate gas exchange properties, the pH is maintained, ATP loss and the release of alpha granular platelet factor 4 (PF4) are prevented. Each of the above references is hereby incorporated in its entirety.

[0009] These findings have led to the standardization of practices to ensure that platelets are oxygenated during room-temperature storage to maximize posttransfusion viability. When platelets are stored at refrigerated temperatures, posttransfusion viability is lost, making such platelets unsuitable for prophylactic transfusion in cancer patients who cannot produce their own platelets. On the other hand, platelets stored at refrigerated temperatures maintain hemostatic function during transfusion. Therefore, viability is less important than hemostatic activity when administering platelets to patients with traumatic bleeding. We have demonstrated that 3 weeks of refrigerated anaerobic storage of whole blood results in hemostatic activity consistent with that of refrigerated conventionally stored whole blood, clearly demonstrating that the hemostatic activity of platelets can be maintained despite hypoxia.

[0010] Although oxygen depletion in whole blood has been documented in the literature, the effects of anaerobic storage of whole blood have not been published. As mentioned above, it is well established that long-term platelet (PLT) survival (greater than 24 hours) requires room temperature storage and oxygen during storage. However, for hemorrhagic trauma resuscitation, long-term PLT survival is less important than their hemostatic potential. Recently, it has become apparent that patients transfused with stored or fresh whole blood, as well as reconstituted whole blood (a mixture of plasma, red blood cells, and platelets), have significantly reduced post-traumatic mortality. We recently discovered that refrigeration allows for anaerobic storage of PLTs and provides the known advantages of anaerobic storage of red blood cells observed in packaged red blood cells. More specifically, deoxygenated whole blood provides improved 2,3-DPG levels, while unexpectedly preserving coagulability without introducing negative effects. During storage, red blood cell deformability is maintained under deoxygenated conditions.

[0011] Oxidative damage during storage has been implicated as a major contributor to damage to the packed red blood cell (pRBC) membrane, as suggested by the accumulation of lipid peroxidation markers such as isoprostanes. Increased cytokine levels during storage may also play a role in the development of storage damage, with potential clinical implications for negative transfusion outcomes.

[0012] Certain patient populations are more susceptible to storage injury than others. Among these more susceptible populations, as non-limiting examples, are trauma patients and cancer patients. Associated with poor clinical outcomes is the accumulation of biological response modifiers (BRMs), which include cytokines that mediate inflammation, regulate cell growth, regulate angiogenesis, and regulate T helper cell function. Among these BRMs are interleukin 17 (IL-17), eotaxin (CCL11), basic FGF (bFGF), macrophage inflammatory protein 1a (MIP-1a), monocyte chemoattractant protein 1 (MCP-1), platelet-derived growth factor (PDGF), tumor necrosis factor alpha (TNF-α), and vascular endothelial growth factor (VEGF). See Behrens et al., "Accumulation of biologic response modifiers during red blood cell cold storage," Transfusion 49 (Supp 13): 10A (2009). Cytokines have also been observed to accumulate during blood storage, and these accumulated cytokines may be associated with poor outcomes when administered perioperatively to cancer patients. See Benson et al., “Accumulation of Pro-Cancer Cytokines in the Plasma Fraction of Stored Packed Red Cells,” J Gastrointest Surg. 16:460-468 (2012). Methods requiring blood storage result in reduced levels of BRMs and cytokines, thereby improving patient outcomes.

[0013] Traumatic injuries account for 30% of all life years lost in the United States, far exceeding cancer (16% of life years) and heart disease (12%). Trauma is the leading cause of death in patients aged 1-46 years. Although hemorrhagic death often occurs within 24 hours of traumatic injury, early death (within 3-6 hours) due to massive hemorrhage is preventable with prompt and appropriate care.

[0014] Damage control resuscitation (DCR) protocols describe the concept of using balanced ratios of blood components. DCR has rapidly become the standard for halting bleeding and reversing shock in acutely bleeding trauma patients. In civilian settings, current blood banking practices do not include whole blood inventory, so DCR employs continuous transfusion of separated blood components (RBCs, plasma, and platelets) to allow the blood to be “reconstituted” in the recipient. Earlier this year, a large randomized controlled trial (RCT), the Pragmatic Randomized Optimizing Platelet and Plasma Ratio (PROPPR), compared the efficacy of transfusing “reconstituted blood” in a 1:1:1 ratio of units (plasma, platelets, and RBCs) to massive transfusion in trauma patients in a 1:1:2 ratio. Massive transfusion kits, which combine prepackaged blood products of thawed fresh frozen plasma (FFP), platelets, and RBCs in a 1:1:1 ratio, are now readily available at major trauma centers.

[0015] Recent studies suggest that whole blood may be superior for controlling bleeding and reversing shock in patients with life-threatening hemorrhage. The 2015 meeting of the NHLBI International Symposium on Transfusion Medicine Research prioritized research on whole blood for patients with severe bleeding. Similarly, the THOR Network, an international organization focused on damage control resuscitation, has compared the efficacy and safety of whole blood with components for hemorrhagic shock. Because modern blood banks do not routinely supply whole blood, over 80% of surveyed level 1 trauma centers attempt to mimic the hemostatic and shock-reversing properties of massive transfusion protocols with a plasma:platelet:red blood cell ratio of 1:1:1 to 1:1:2 for both traumatic and nontraumatic life-threatening hemorrhage. Logistically, providing all three blood components quickly and safely is challenging, especially given the need to thaw plasma in centers that do not have immediate access to a thawed plasma inventory. Recent data also suggest that whole blood stored at 4°C for up to 14 days maintains platelet function and global hemostatic efficacy, surpassing storage at 22°C.

[0016] In addition to requiring blood banks to provide whole blood for certain patient populations, the ability to preserve valuable blood resources is also important. Particularly, blood banks typically discard whole blood inventory (even if FDA regulations allow longer service life) after 2 weeks, so valuable and often scarce resources are not utilized. This ability to maximize the value of blood resources is particularly useful for small hospitals serving as III and IV level trauma centers, where the hemostatic whole blood product of anoxic conditions can be maintained under anaerobic conditions and then packaged for red blood cell processing. This specification sheet provides improved whole blood quality for trauma patients, and further provides another packaged red blood cell source with improved properties and reduced storage damage. This specification sheet overcomes the worry about consuming valuable O-type negative RBCS (generally used for whole blood transfusion). Therefore, anaerobic red blood cells can be obtained from the whole blood that oxygen reduces and are recycled into the red blood cell unit of anoxic conditions, suitable for storage for up to six weeks. As provided herein, the red blood cells of deoxygenated packaging can be obtained from unused oxygenated whole blood, for blood transfusion, or stored for use later under anaerobic conditions. Summary of the Invention

[0017] The present disclosure provides and encompasses methods for improving the survival of patients requiring multiple transfusions comprising providing stored red blood cells that have had their oxygen reduced to a patient in need thereof undergoing a medical procedure.

[0018] The present disclosure provides and encompasses methods for improving survival of a cancer patient in need thereof following a perioperative transfusion, the methods comprising providing oxygen-reduced stored red blood cells to a cancer patient requiring a surgical procedure.

[0019] The present disclosure provides and includes a method for reducing cancer-promoting factors in stored blood, comprising depleting oxygen from the blood prior to storage, the method comprising collecting blood in an anticoagulant solution, reducing white blood cells from the collected blood, reducing the oxygen saturation (SO2) to below 30% before storage, reducing the partial pressure of carbon dioxide to below 60 mmHg before storage, and storing the oxygen and carbon dioxide reduced blood under anaerobic conditions.

[0020] The present disclosure provides and includes a blood composition for transfusion into a trauma patient in need thereof, comprising deoxygenated leukocyte-reduced whole blood in an anticoagulant solution and having a pre-stored oxygen saturation (SO2) of 20% or less and a pre-stored partial pressure of carbon dioxide of less than 60 mmHg, wherein the deoxygenated leukocyte-reduced whole blood has a 2,3-DPG level at 15 days that is higher than the initial 2,3-DPG level of the deoxygenated leukocyte-reduced blood.

[0021] The present disclosure provides and encompasses methods of reducing an inflammatory response in a patient receiving a blood transfusion, the methods comprising delivering an oxygen-depleted blood product to a patient in need thereof, wherein the oxygen-depleted whole blood has reduced levels of inflammatory cytokines after storage under anaerobic conditions.

[0022] The present disclosure provides and includes methods for reducing an immune response in a patient receiving a blood transfusion, the methods comprising delivering an oxygen-depleted blood product to a patient in need thereof, wherein the oxygen-depleted blood product has reduced cytokine levels after storage under anaerobic conditions. In aspects according to the present disclosure, the immune response is immunomodulatory or immunosuppressive. In other aspects, the immune response is activation, including, for example, inflammation.

[0023] The present disclosure provides and includes methods for improving oxygen perfusion in a patient in need thereof, the methods comprising delivering an oxygen-depleted blood product to a patient in need thereof, wherein the oxygen-depleted blood product has a higher RBC deformability than conventionally stored blood products.

[0024] The present disclosure provides and includes a method for managing a blood bank, comprising: maintaining blood units comprising oxygen-reduced whole blood and an anticoagulant or oxygen-reduced leukopenic whole blood and an anticoagulant; providing one or more blood units from the inventory for use in treating a patient; and recovering blood units from the inventory to prepare fractionated, deoxygenated blood units. The present disclosure further provides for using recycled blood units to prepare regenerated blood units for use in the treatment of trauma patients requiring massive transfusions.

[0025] The present disclosure provides and includes methods of providing a supply of blood products for use in transfusion medicine, comprising depleting oxygen or oxygen and carbon dioxide from whole blood to prepare oxygenated or oxygen- and carbon dioxide-reduced whole blood; and storing the oxygenated or oxygen- and carbon dioxide-reduced whole blood for a period of time and providing the stored blood to a patient in need thereof; or storing the oxygenated or oxygen- and carbon dioxide-reduced whole blood for a period of time and preparing oxygenated or oxygen- and carbon dioxide-reduced packaged red blood cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The present disclosure is disclosed with reference to the accompanying drawings, in which:

[0027] Figures 1A to 1D are graphs showing the results of cytokine measurements, demonstrating reduced levels of eotaxin (1A) and RANTES (1B) in anaerobically stored packaged erythrocytes. Figure 1C Shows reduced cell-free hemoglobin levels compared to aerobically stored packed red blood cells. Figure 1D Shown is the reduced isoprostane content in anaerobically stored packed red blood cells. Dashed line = aerobically stored blood; solid line = anaerobically stored blood.

[0028] Figures 2A to 2G is a graph showing the results of two experiments according to the present invention comparing storage of leukocyte-reduced whole blood in anticoagulant solution CPD (LRWB / CPD) collected over 21 days with oxygen-reduced, oxygen- and carbon dioxide-reduced, and conventionally stored LRWB / CPD. Figure 2A 2,3-DPG levels are shown. Figure 2B ATP levels are shown. Figure 2C The pH is shown. Figure 2D Displays the platelet count. Figure 2E Present potassium levels. Figure 2F Presentation Figure 2A Data replotted relative to 2,3-DPG levels on day 0 (T0). Figure 2G show Figure 2B Data are replotted against ATP levels on day 0 (T0). Key: Sample c68 / 80 is conventionally stored blood with an initial oxygen saturation of 68% and a partial pressure of carbon dioxide of 80 mmHg; sample c50 / 94 is conventionally stored blood with an initial oxygen saturation of 50% and a partial pressure of CO2 of 94 mmHg; sample sc91 / 75 is conventionally stored blood with an initial oxygen saturation of 91% and a partial pressure of CO2 of 75 mmHg; sample sc69 / 87 is conventionally stored blood with an initial oxygen saturation of 69% and a partial pressure of CO2 of 87 mmHg; sample tc5 / 78 The blood was oxygen-depleted, with an anaerobically stored blood having an initial oxygen saturation of 5% and a CO2 partial pressure of 78 mmHg; sample tc7 / 64 was oxygen-depleted, with an anaerobically stored blood having an initial oxygen saturation of 7% and a CO2 partial pressure of 64 mmHg; sample T5 / 28 was oxygen- and carbon dioxide-depleted, with an anaerobically stored blood having an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; sample T4 / 26 was oxygen- and carbon dioxide-depleted, with anaerobically stored blood having an initial oxygen saturation of 4% and a CO2 partial pressure of 26 mmHg.

[0029] Figures 3A to 3D is a graph showing the results of two experiments according to the present disclosure comparing the anticoagulant solution CPDA1 (LRWB / CPDA1) in oxygen-reduced, oxygen- and carbon dioxide-reduced, and conventionally stored LRWB / CPDA1 in leukoreduced whole blood collected over 21 days. Figure 3A 2,3-DPG levels are shown. Figure 3B ATP levels are shown. Figure 3C Shown are replotted relative to the 2,3-DPG level on day 0 (T0). Figure 3A data. Figure 3D The ATP levels at day 0 (T0) are re-plotted. Figure 3BKey: Sample C32 / 98 was conventionally stored blood with an initial oxygen saturation of 32% and a CO2 partial pressure of 98 mmHg; Sample C56 / 86 was conventionally stored blood with an initial oxygen saturation of 56% and a CO2 partial pressure of 86 mmHg; Sample SC59 / 95 was conventionally stored blood with an initial oxygen saturation of 59% and a CO2 partial pressure of 95 mmHg; Sample SC82 / 84 was conventionally stored blood with an initial oxygen saturation of 82% and a CO2 partial pressure of 84 mmHg; Sample TC7 / 80 .... The blood was oxygen-depleted, with an anaerobically stored blood having an initial oxygen saturation of 7% and a CO2 partial pressure of 80 mmHg; sample TC6 / 77 was oxygen-depleted, with an anaerobically stored blood having an initial oxygen saturation of 6% and a CO2 partial pressure of 77 mmHg; sample T5 / 28 was oxygen- and carbon dioxide-depleted, with an anaerobically stored blood having an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; and sample T7 / 23 was oxygen- and carbon dioxide-depleted, with anaerobically stored blood having an initial oxygen saturation of 7% and a CO2 partial pressure of 23 mmHg.

[0030] Figures 4A to 4C is a graph showing experimental results according to the present disclosure comparing leukocyte-reduced whole blood collected in anticoagulant solution CPDA1 (LRWB / CPDA1) during a 21-day period under oxygen-reduced (OR), oxygen and carbon dioxide-reduced (OCR), and conventionally stored LRWB / CPDA1. Figure 4A The levels of ATP in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1 are shown. Figure 4B The levels of 2,3-DPG in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1 are shown. Figure 4C The percentage of hemolysis in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1 is shown. Figures 4A to 4C In the graph presented in , small dashed line = OR-LRWB / CPDA1 stored blood, dashed line = ORC-LRWB / CPDA1 stored blood, solid line = conventionally stored blood.

[0031] Figures 5A to 5D is a graph showing experimental results according to the present invention comparing storage of leukocyte-reduced whole blood in anticoagulant solution CPDA1 (LRWB / CPDA1) collected over 21 days with reduced oxygen and carbon dioxide and conventionally stored LRWB / CPDA1. Figure 5A Shown are the activated partial thromboplastin times (seconds) (aPTT) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5B Prothrombin time (seconds) (PT) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 is shown. Figure 5C Fibrinogen levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 are shown. Figure 5D D-dimer levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 are shown. In the graphs showing plasma coagulation parameters, dashed line = ORC-LRWB / CPDA1 stored blood, solid line = conventionally stored blood.

[0032] Figures 6A to 6E is a graph showing experimental results according to the present invention comparing storage of leukocyte-reduced whole blood in anticoagulant solution CPDA1 (LRWB / CPDA1) collected over 21 days with reduced oxygen and carbon dioxide and conventionally stored LRWB / CPDA1. Figure 6A Factor V levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 are shown. Figure 6B Factor VIII levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 are shown. Figure 6C Protein C activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 is shown. Figure 6D Protein S activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 is presented. Figure 6E The levels of von Willebrand Factor (vWF) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 are shown. In the graphs showing plasma coagulation factors, dotted line = ORC-LRWB / CPDA1 stored blood, solid line = conventionally stored blood.

[0033] 7A to 7D is a graph showing experimental results according to the present invention comparing leukocyte-reduced whole blood in oxygen and carbon dioxide reduced (OCR) and conventionally stored LRWB / CPDA1 collected over 21 days in anticoagulant solution CPDA1 (LRWB / CPDA1). Figure 7A Shown are the rates of fibrin aggregation and cross-linking in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 (TEG perspective). Figure 7B Comparison of hemodynamics (TEG K) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 is shown. Figure 7C The maximum amplitudes in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1 are given. Figure 7DReaction times are shown for OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graphs presenting thromboelastography (TEG) parameters, dashed line = ORC-LRWB / CPDA1 stored blood, solid line = conventionally stored blood. DETAILED DESCRIPTION

[0034] Unless otherwise defined, the technical and scientific terms used herein have the same meanings as those generally understood by those of ordinary skill in the art. Those skilled in the art will recognize that many methods can be used in the practice of the present disclosure. In fact, the present disclosure is in no way limited to the methods and materials described. Any references cited herein are incorporated by reference in their entirety. For the purposes of this disclosure, the following terms are defined as follows.

[0035] As used herein, the term "patient" includes a person undergoing a medical procedure requiring the receipt of a blood product.

[0036] As used herein, the term "multiple transfusions" includes patients who have received more than 195 units of blood. In another aspect, multiple transfusions may include patients who have received at least 1×10 5 mL of blood. On the other hand, multiple transfusions involve receiving 1 to 1 × 10 5 mL of blood. On the other hand, multiple transfusions involve receiving 1×10 4 to 1×10 5 mL of blood in patients.

[0037] As used herein, the term "blood" refers to whole blood, leukopenic RBCs, thrombocytopenic RBCs, and leukocyte and thrombocytopenic RBCs. The term blood also includes packed red blood cells, thrombocytopenic packed red blood cells, leukocyte-reduced packed red blood cells (LRpRBCs), and leukocyte and thrombocytopenic packed red blood cells. The temperature of the blood can start from the normal body temperature of 37°C at the time of collection, depending on the stage of the collection process, and once the blood leaves the patient's body, it immediately drops to about 30°C, and then after about 6 hours, it further drops to room temperature, and finally is refrigerated between about 4°C and 6°C.

[0038] As used herein, "blood products" include separated platelets, plasma, or white blood cells.

[0039] As used herein, "recovered blood product" includes separated platelets, plasma, or white blood cells collected from a donor.

[0040] As used herein, " recovered blood " includes whole blood and red blood cells collected from a donor and stored in advance under conditions of oxygen reduction. In one aspect of the present disclosure, suitable blood for the method includes packaged red blood cells (OR-LRpRBC) reduced in oxygen and leukocytes, packaged red blood cells (OR-LRpRBC+PLT) reduced in oxygen and leukocytes and comprising platelets, packaged red blood cells (OCR-LRpRBC) reduced in oxygen and carbon dioxide, or packaged red blood cells (OCR-LRpRBC) reduced in oxygen and carbon dioxide, and the platelets obtained from oxygen-reduced leukocyte-reduced whole blood (OR-LRWB) are reduced, the platelets are reduced and the whole blood (OR-LRWB+PLT) is reduced in oxygen and carbon dioxide, and the platelets are reduced and the whole blood (OCR-LRWB+PLT) is reduced. On the other hand, suitable blood storage for the method is up to 42 days. In another aspect, suitable blood for use in the method is stored for up to 56 days. In another aspect, suitable blood for use in the method is stored for up to 64 days.

[0041] As used herein, a method of obtaining a "fractionated blood product" comprises obtaining recycled blood from a blood bank inventory and separating it into platelets, plasma, and leukocytes. Suitable blood for use in this method includes oxygenated whole blood with an anticoagulant and oxygen-reduced leukocyte-reduced whole blood with an anticoagulant. In one aspect of the present disclosure, the separated components of the oxygen-reduced blood are stored for up to six weeks. In another aspect, separating the components of the oxygen-reduced blood includes an additive solution. In certain aspects, the additive solution can be AS-1. In certain aspects, the additive solution is AS-3 ( ). In some aspects, the additive solution is AS-5. In some aspects, the additive solution is SAGM. In some aspects, the additive solution is PAGG-SM. In some aspects, the additive solution is PAGG-GM. In some aspects, the additive solution is MAP. In some aspects, the additive solution is SOLX. In some aspects, the additive solution is ESOL. In some aspects, the additive solution is EAS61. In some aspects, the additive solution is OFAS1. In some aspects, the additive solution is OFAS3. In some aspects, the additive solution is AS-1, AS-3 ( ), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, SOLX, ESOL, EAS61, OFAS1 and OFAS3, used alone or in combination.

[0042] As used herein, "reconstituted WB" includes providing platelets, RBCs, and plasma in parallel with the patient during a transfusion.

[0043] As used herein, "derived WB" includes oxygen-reduced as well as oxygen- and carbon dioxide-reduced whole blood.

[0044] As used herein, "stored red blood cells" include red blood cells that are stored at 1 to 6°C in oxygen-reduced or oxygen- and carbon dioxide-reduced red blood cells. In one aspect, stored red blood cells include red blood cells (RBCs) present in whole blood. In another aspect, stored red blood cells include red blood cells present in leukopenic whole blood. In another aspect, stored red blood cells include red blood cells (RBCs) present in leukopenic RBCs. In another aspect, stored red blood cells include red blood cells (RBCs) present in thrombocytopenic RBCs. In another aspect, stored red blood cells include red blood cells (RBCs) present in leukopenic and thrombocytopenic red blood cells.

[0045] As used herein, “whole blood” includes white blood cells (WBC), platelets suspended in plasma, and also includes electrolytes, hormones, vitamins, antibodies, etc. In whole blood, the number of white blood cells is usually between 4.5 and 11.0×10 9 Normal RBC levels range from 4.6 to 6.2 × 10 cells / L for men. 12 / L, and 4.2-5.4×10 12 / L. A normal hematocrit, or percentage packed cell volume, is approximately 40-54% for men and 38-47% for women. Platelet counts are typically 150-450 × 10 9 / L. Whole blood is collected from a blood donor, typically in combination with an anticoagulant. Whole blood is initially collected at about 37°C and rapidly cooled to about 30°C during and shortly after collection, but slowly cooled to ambient temperature over about 6 hours. Whole blood can be collected according to the methods of the present disclosure starting at 30-37°C, or at room temperature (typically about 25°C). As used herein, a "unit" of blood is about 450-500ml, including anticoagulant. Suitable anticoagulants include CPD, CPDA1, ACD, and ACD-A. As used herein, "time of collection" (Tc) is the time at which blood is collected from a patient.

[0046] As used herein, "red blood cells" (RBC), stored red blood cells, oxygen-reduced red blood cells, oxygen- and carbon dioxide-reduced red blood cells include red blood cells in whole blood, leukopenic RBC, thrombocytopenic RBC, leukocytes, thrombocytopenic RBC, and packaged red blood cells (pRBC). Red blood cells are in a dynamic state in the human body. Red blood cells contain hemoglobin, which is an iron-containing protein that carries oxygen to the entire body and makes the blood red. The percentage of blood volume composed of red blood cells is called hematocrit. As used herein, unless otherwise specified, RBC also includes packaged red blood cells (pRBC). Packaged red blood cells are prepared from whole blood using centrifugation techniques known in the art. As used herein, unless otherwise noted, the hematocrit of pRBC is about 70%. As used herein, oxygen-reduced RBC (OR-RBC) can include oxygen- and carbon dioxide (OCR-)-reduced RBC (OCR-RBC).

[0047] As used herein, "leukoreduced whole blood" (LRWB) includes anticoagulant whole blood that has been processed to remove white blood cells and platelets, typically by filtration or centrifugation. Leukoreduced whole blood has at least a 5 log reduction in white blood cell levels.

[0048] As used herein, "oxygen reduced leukopenic whole blood" (OR-LRWB) may include oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB).

[0049] As used herein, "leukocyte-reduced whole blood containing platelets" (LRWB+PLT) includes oxygen-reduced (OR-) whole blood with an anticoagulant and a platelet-conserving filter. As used herein, oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT) may include oxygen- and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT).

[0050] As used herein, "leukopenia packaged red blood cells" (LRpRBC) include packaged red blood cells with oxygen-reduced (OR-) whole blood that has an anticoagulant that has been filtered or centrifuged to remove white blood cells. As used herein, oxygen-reduced leukopenia packaged red blood cells (OR-LRpRBC) may include oxygen- and carbon dioxide-reduced leukopenia packaged red blood cells (OCR-LRpRBC).

[0051] As used herein, "leukopenia-containing packaged red blood cells" (LRpRBC+PLT) include packaged red blood cells with platelets obtained from oxygen-reduced whole blood, which have an anticoagulant that has been passed through a platelet protection filter to remove white blood cells. As used herein, oxygen-reduced leukopenia-containing packaged red blood cells (OR-LRpRBC+PLT) may include oxygen- and carbon dioxide-reduced leukopenia-containing packaged red blood cells (OCR-LRpRBC+PLT).

[0052] In various aspects of the present disclosure, the methods and compositions can include adding an additive solution to the packaged RBCs to form a suspension. Many additive solutions are known in the art. In certain aspects, the additive solution can be selected from the group consisting of AS-1, AS-3 ( ), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5, EAS61, OFAS1 and OFAS3, used alone or in combination. Additive AS-1 is disclosed in "Use of Adsol preservation solution for prolonged storage of low viscosity AS-1 red blood cells," Br J Haema tol., 57(3): 467-78 (1984) by Heaton et al. In another aspect, the additive solution can have a pH of 5.0 to 9.0. In another aspect, the additive can include an antioxidant. In some aspects according to the present disclosure, the antioxidant can be quercetin, α-tocopherol, ascorbic acid, or an enzyme inhibitor of an oxidase.

[0053] As used herein, the term "about" refers to ±10%.

[0054] The terms "include," "comprising," "containing," "having" and their conjugations mean "including but not limited to."

[0055] The term "consisting of" means "including and limited to."

[0056] The term "consisting essentially of" means that the composition, method or structure may include additional ingredients, steps and / or parts, but only if the additional ingredients, steps and / or parts do not materially change the basic and novel characteristics of the claimed composition, method or structure.

[0057] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof.

[0058] Throughout this application, various aspects of the present disclosure can be presented in the form of ranges. It should be understood that the description of range form is merely for convenience and brevity, and should not be interpreted as an inflexible limitation on the scope of the present disclosure. Therefore, the description of a range should be considered to specifically disclose all possible subranges and individual numerical values ​​within the range. For example, the description of a range such as "1 to 6" should be considered to have specifically disclosed subranges, such as "1 to 3," "1 to 4," "1 to 5," "2 to 4," "2 to 6," "3 to 6," etc., and individual numbers within the range, such as 1, 2, 3, 4, 5, and 6. This applies regardless of the width of the scope.

[0059] Whenever a numerical range is indicated herein, it is intended to include any cited numeral (fractional or integer) within the indicated range. "A range" between a first indicated numeral and a second indicated numeral and "a range from" a first indicated numeral" to" a second indicated numeral" are used interchangeably herein and are intended to include the first and second indicated numerals and all fractional and integer numerals therebetween.

[0060] As used herein, the term "method" refers to manners, means, techniques and procedures for accomplishing a given task, including but not limited to those manners, means, techniques and procedures known to those skilled in the art of chemistry, pharmacology, biology, biochemistry and medicine or readily developed from known manners, means, techniques and procedures.

[0061] As used herein, the term "equivalent" means that the measured values ​​of oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are within 1 standard deviation of each other under each compared measurement condition when compared to the measured values ​​of otherwise equivalently treated conventionally stored blood, with a sample size of at least 5.

[0062] As used herein, the term "greater" or "increased" refers to a measurement of oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) that is at least 1 standard deviation greater when compared to the measurement of otherwise equivalently treated conventionally stored blood when compared to OR-WB, with a sample size of at least 5 for each compared measurement condition.

[0063] As used herein, the term "reduced" or "reduced" means that the measured value of oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is at least 1 standard deviation lower when compared to the measured value of otherwise equivalently treated conventionally stored blood when compared to OR-WB, with a sample size of at least 5 for each compared measurement condition.

[0064] As used herein, the terms "conventional storage," "conventional storage," and "conventional conditions" include whole blood, leukopenic RBCs, thrombocytopenic RBCs, leukocytes, thrombocytopenic RBCs, packaged red blood cells, thrombocytopenic packaged red blood cells, leukocyte-reduced packaged red blood cells (LRpRBCs), leukocytes, and thrombocytopenic packaged red blood cells stored in oxygen and carbon dioxide permeable containers without a gas reduction step at 1 to 6°C prior to storage. In one aspect of the present disclosure, due to the permeability of the oxygen and carbon dioxide containers, oxygen and carbon dioxide increase to ambient levels over time in conventionally stored whole blood, leukopenic RBCs, thrombocytopenic RBCs, leukocytes, and thrombocytopenic RBCs, packaged red blood cells, thrombocytopenic packaged red blood cells, leukocyte-reduced packaged red blood cells (LRpRBCs), leukocytes, and thrombocytopenic packaged red blood cells. Although not conventionally considered conventional, for the purposes of this disclosure, conventional storage may include storage at temperatures above 6°C. Furthermore, although not traditionally considered conventional, for purposes of this disclosure, conventional storage may include storage at refrigerated temperatures.

[0065] The present disclosure provides and includes methods for providing desirable characteristics of blood products for transfusion. It has been found that depletion of oxygen from packaged red blood cells results in a decrease in the accumulation of unbound cytokines, particularly RANTES (CC motif chemokine ligand 5, CCL5) and eotaxin (CC motif chemokine ligand 11, CCL11), as well as cell-free hemoglobin and 8-isoprostane F. 2αWithout being limited by theory, it is believed that RANTES and eosinophil chemokine are normally sequestered by binding to DARC (atypical chemokine receptor 1, ACKR1), and that oxidative stress damages DARC and releases the bound chemokines. Therefore, although the total content of chemokines remains unchanged, the effective concentration (e.g., freely diffusing and unbound) increases and can be used to influence transfused patients. As will be appreciated, the presence of these active chemokines (which act in a dose-dependent manner) may be harmful to trauma and other patients who have received two or more transfusions. In addition to the required effective oxygen delivery associated with elevated 2,3-DPG values, these findings indicate unexpected benefits of anaerobically stored blood and provide a potential reduction in some components of storage damage resulting from pRBC oxidative damage during storage. These cytokines are known to be negatively correlated with patient outcomes in some patient populations. Therefore, the discovery that unbound cytokine accumulation can be reduced provides an improved method for treating patients who are sensitive to cytokines.

[0066] The present disclosure provides and includes improving the survival of patients who require multiple blood transfusions by providing stored red blood cells that have undergone oxygen reduction (OR-stored RBCs) to patients who need to undergo medical procedures. Without being limited by theory, it is believed that increased cytokine levels have an adverse effect on the recipient patient, increasing morbidity. In one aspect, the stored red blood cells are oxygen reduced (OR). In another aspect, the stored red blood cells are both oxygen and carbon dioxide reduced (OCR). As shown in the Examples, in OCR samples, ATP levels are reduced and maintained at a lower level for at least 15 days, while in OR samples, ATP levels are increased compared to conventionally stored samples (see Figure 4A As shown in the Examples, in the OCR samples, the level of 2,3-DPG increased and remained at a high level for at least 15 days, while in the OR samples, the level of 2,3-DPG increased compared to conventional storage, but was not higher than that of the OCR samples (see Figure 4B ). Furthermore, as shown in the Examples, hemolysis was comparable in OR, OCR, and conventionally stored samples.

[0067] In one aspect of the present disclosure, the cytokine comprises monocyte chemoattractant protein-1 (MCP-1). In another aspect, the cytokine comprises RANTES, which regulates normal T-cell expression and secretion on activated cells. In another aspect, the cytokine comprises angiogenin. In another aspect of the present disclosure, the cytokine comprises tumor necrosis factor-α (TNF-α). In another aspect, the cytokine comprises epidermal growth factor (EGF). In another aspect, the cytokine comprises platelet-derived growth factor (PDGF).

[0068] In one aspect of the present disclosure, after 21 days under OR conditions, the level of factor RANTES is less than 500 pg / ml. In another aspect, after 21 days under OR conditions, the level of factor RANTES is less than 400 pg / ml. In another aspect, after 21 days under OR conditions, the level of factor RANTES is less than 300 pg / ml. In another aspect, after 21 days under OR conditions, the level of factor RANTES is greater than 100 pg / ml. In another aspect, after 21 days under OR conditions, the level of factor RANTES is between 0 and 300 pg / ml.

[0069] In one aspect of the present disclosure, after 21 days under OR conditions, the level of eotaxin is less than 150 pg / ml. In another aspect, after 21 days under OR conditions, the level of eotaxin is less than 100 pg / ml. In another aspect, after 21 days under OR conditions, the level of eotaxin is between 0 and 100 pg / ml. In another aspect, after 21 days under OR conditions, the level of eotaxin is preferably 100 pg / ml. In another aspect, after 21 days under OR conditions, the level of eotaxin is greater than 100 pg / ml. In another aspect, after 21 days under OR conditions, the level of eotaxin is between 0 and 300 pg / ml.

[0070] In aspects according to the present disclosure, the OR-stored RBCs are selected from: oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen-reduced leukocyte-reduced packaged red blood cells (OR-LRpRBC), oxygen-reduced leukocyte-reduced packaged red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), oxygen and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), oxygen and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), and combinations thereof. In other aspects, the OR-stored RBCs comprise oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT).

[0071] In one aspect, the patient requiring multiple transfusions is a trauma patient. In another aspect, the patient requiring multiple transfusions is a transplant patient. In another aspect, the patient requiring multiple transfusions is a cardiac surgery patient. In another aspect, the patient requiring multiple transfusions is an obstetric patient. In another aspect, the patient requiring multiple transfusions is a gastrointestinal (GI) surgery patient. In another aspect, the patient is an orthopedic surgery patient.

[0072] On one hand, patients who require multiple blood transfusions are trauma patients. On the other hand, patients who require multiple blood transfusions are hemorrhagic trauma patients. On the other hand, patients who require multiple blood transfusions are blunt trauma patients.

[0073] In one aspect, reducing cytokines in oxygen-depleted stored red blood cells provides improved treatment for cancer patients requiring blood transfusions. It is known in the art that cytokines are negatively correlated with patient outcomes in surgical treatment of cancer patients who receive perioperative blood transfusions. In one aspect, an oxygen-depleted, cytokine-reduced blood product is provided to a cancer patient prior to surgery. In another aspect, an oxygen-depleted, cytokine-reduced blood product is provided to a cancer patient during surgery. In another aspect, an oxygen-depleted, cytokine-reduced blood product is provided to a cancer patient postoperatively.

[0074] In aspects according to the present disclosure, oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have higher levels of 2,3-DPG than traditionally stored leukopenic whole blood (WB) and provide improved oxygen delivery. Under anaerobic conditions, 2,3-DPG levels can be maintained in whole blood for up to 4 weeks. In one aspect, 2,3-DPG levels are maintained at more than 50% of physiological levels for up to four weeks. In aspects according to the present disclosure, improved 2,3-DPG levels are maintained at 2 weeks. In other aspects, 2,3-DPG levels are maintained for three weeks. In one aspect, the level of 2,3-DPG in oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) is at least 80% or more higher than the 2,3-DPG level in blood on day 0. In another aspect, the level of 2,3-DPG in oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) is at least 5 to 20 DPG μmol / gHb.

[0075] Also provided and included in the present disclosure are: oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT). Platelet-depleted whole blood (OCR-LRWB+PLT) has been depleted of whole blood (OR-WB) and has reduced levels of biological response modifiers (BRMs) relative to conventionally stored whole blood. In certain aspects, the BRMs present in oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) are about half the amount of conventionally stored blood after 21 days. On the one hand, oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have cytokine levels that are relatively unchanged after storage for 10 days under anaerobic conditions. On the other hand, cytokine levels are relatively unchanged after storage for 30 days. On the other hand, cytokine levels are relatively unchanged after storage for 40 days. As used herein, "relatively unchanged" means that the concentration of the cytokine and the hemoglobin level are normalized to within 1 standard deviation of the initial standardized concentration of the cytokine.

[0076] In some aspects, compared with the whole blood of traditional storage, oxygen reduces the whole blood (OR-LRWB) that leukopenia is reduced, oxygen reduces the whole blood (OR-LRWB+PLT) that leukopenia is reduced and comprises platelet, oxygen and carbon dioxide reduce the whole blood (OCR-LRWB), oxygen and carbon dioxide reduce the whole blood (OCR-LRWB) that leukopenia is reduced and comprises platelet, and the eotaxin of the cytokine level of reduction is reduced.In some aspects, the level of eotaxin in oxygen reduces the whole blood (OR-LRWB), oxygen reduces the whole blood (OR-LRWB+PLT) that leukopenia is reduced and comprises platelet, oxygen and carbon dioxide reduce the whole blood (OCR-LRWB), oxygen and carbon dioxide reduce the whole blood (OCR-LRWB) that leukopenia is reduced and comprises platelet, and the eotaxin of the whole blood (OCR-LRWB+PLT) that leukopenia is reduced is about half of the level of eotaxin of traditional storage blood after 21 days hemoglobin concentration normalization. In one aspect, the level of eotaxin in oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is about 25% or less of the level of eotaxin in conventionally stored blood after 40 days.

[0077] In certain aspects, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) have reduced levels of the cytokine RANTES (regulated activation, normal T cell expressed and secreted) compared to conventionally stored whole blood. In one aspect, the level of RANTES in oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB+PLT) is approximately half the level of RANTES present in conventionally stored blood after 21 days of normalization of hemoglobin concentration. In one aspect, the level of RANTES in oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is about 25% or less of the level of RANTES present in conventionally stored blood after 40 days.

[0078] In certain aspects, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood comprising platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood comprising platelets (OCR-LRWB+PLT) have reduced monocyte chemoattractant protein-1 (MCP-1) levels compared to traditional stored whole blood. On the one hand, the level of MCP-1 in oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood comprising platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB+PLT) is about half the level of MCP-1 that exists in traditional stored blood after 21 days of normalization of hemoglobin concentration. In one aspect, the level of MCP-1 in oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is approximately 25% or less of the level of MCP-1 present in conventionally stored blood after 40 days.

[0079] In certain aspects, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) have reduced levels of angiogenin compared to conventionally stored whole blood. In one aspect, the level of angiogenin in oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB+PLT) is about half the level of angiogenin present in conventionally stored blood after 21 days of normalization of hemoglobin concentration. In one aspect, the level of angiogenin in oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is approximately 25% or less of the level of angiogenin present in conventionally stored blood after 40 days.

[0080] In certain aspects, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) have reduced levels of tumor necrosis factor-alpha (TNF-alpha) compared to conventionally stored whole blood. In one aspect, the level of TNF-alpha in oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) is approximately half the level of TNF-alpha present in conventionally stored blood after 21 days of normalization of hemoglobin concentration. In one aspect, the level of TNF-α in oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is about 25% or less of the level of TNF-α present in conventionally stored blood after 40 days.

[0081] In certain aspects, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) have reduced levels of epidermal growth factor (EGF) compared to traditionally stored whole blood. In one aspect, the level of EGF in oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) is about half the level of EGF present in traditionally stored blood after 21 days of normalization of hemoglobin concentration. In one aspect, the level of EGF in oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is about 25% or less of the level of EGF present in conventionally stored blood after 40 days.

[0082] In certain aspects, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) have reduced levels of soluble CD40 ligand (sCD40L) compared to conventionally stored whole blood. In one aspect, the level of sCD40L in oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) is approximately half the level of sCD40L present in conventionally stored blood after 21 days of normalization of hemoglobin concentration. In one aspect, the level of sCD40L in oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is about 25% or less of the level of sCD40L present in conventionally stored blood after 40 days.

[0083] In certain aspects, oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have reduced levels of platelet-derived growth factor (PDGF) compared to conventionally stored whole blood. In one aspect, the level of PDGF in oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), is about half the level of PDGF present in conventionally stored blood after 21 days of normalization of hemoglobin concentration. In one aspect, the level of PDGF in oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is about 25% or less of the level of PDGF present in conventionally stored blood after 40 days.

[0084] The present disclosure provides and includes oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), which provide a reduced inflammatory response when transfused into a patient, compared to conventionally stored oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT).

[0085] The present disclosure provides and includes oxygen-reduced leukopenic whole blood (OR-LRWB) that provides a blood product having a higher RBC deformability than conventional stored blood products. In certain aspects, the blood product is a whole blood product. In another aspect, the blood product is leukopenic whole blood. In another aspect, the blood product is leukopenic and thrombocytopenic whole blood. In another aspect, the blood product is leukopenic packaged red blood cells or leukocyte and platelet-reduced packaged red blood cells.

[0086] The present disclosure provides and includes oxygen-reduced leukopenia whole blood (OR-LRWB) having coagulation parameters that are at least 75% of the coagulation parameters of traditional stored whole blood measured by thromboelastography (TEG). On the one hand, the TEG coagulation parameters of oxygen-reduced leukopenia whole blood (OR-LRWB), oxygen-reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenia whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) are equivalent to traditional stored blood. In another aspect, oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have TEG coagulation parameters that are greater than TEG coagulation parameters of conventionally stored blood. In one aspect, the TEG angle is greater than 40°. In another aspect, the TEG kinetics (K) is less than 5 minutes. In another aspect, the TEG K is between 1 and 5 minutes. In another aspect, the TEG maximum amplitude (TEG MA) is greater than 50 mm. In another aspect, the TEG maximum amplitude (TEG MA) is less than 70 mm. In another aspect, the TEG maximum amplitude (TEG MA) is between 30 and 65 mm. In another aspect, the TEG reaction time (TEGR) is less than 10 minutes. In another aspect, the TEG reaction time (TEG R) is less than 8 minutes. In another aspect, the TEG reaction time (TEG R) is at least 3 minutes. On the other hand, the TEG reaction time (TEGR) is between 4 and 8 minutes.

[0087] The present disclosure provides oxygen-reduced leukocyte-reduced whole blood (OR-LRWB) having coagulation parameters that are at least 75% of the coagulation parameters of traditional stored whole blood measured by prothrombin time (PT). On the one hand, the PT of oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are equivalent to traditional stored blood. On the other hand, oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have a PT greater than the PT of traditional stored blood. In another aspect, the oxygen reduced leukocyte reduced whole blood (OR-LRWB) has a PT of less than 15 seconds. In another aspect, the oxygen reduced leukocyte reduced whole blood (OR-LRWB) has a PT of greater than 5 seconds. In another aspect, the oxygen reduced leukocyte reduced whole blood (OR-LRWB) has a PT of 10 to 15 seconds.

[0088] The present disclosure provides and includes oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), whose coagulation parameters are at least 75% of the coagulation parameters of traditionally stored whole blood (measured by partial thromboplastin time (PTT)). In one aspect, the PTT of oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is equivalent to that of traditionally stored blood. In yet another aspect, oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have a PTT greater than the PTT of conventionally stored blood. In another aspect, oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have a PTT greater than 25 seconds. On the other hand, oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have a PTT of less than 40 seconds. On the other hand, oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have a PTT between 32 and 42 seconds.

[0089] The present disclosure provides and includes oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), having whole blood (OR-WB) with coagulation parameters that are at least 75% of the coagulation parameters of conventionally stored whole blood, as measured by fibrinogen activity levels. In one aspect, the fibrinogen activity of oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is equivalent to that of conventionally stored blood. In yet another aspect, oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have a fibrinogen activity greater than the fibrinogen activity of conventionally stored blood. In another aspect, oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have a fibrinogen level of at least 200 mg / ml. On the other hand, oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have a fibrinogen level of up to 400 mg / ml. On the other hand, oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have a fibrinogen level of 250 to 350 mg / ml.On the other hand, oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have fibrinogen levels of 250 to 300 mg / ml.

[0090] The present disclosure provides and includes oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) having coagulation parameters as measured by D-dimer analysis that are at least 75% of the coagulation parameters of conventionally stored whole blood (OR-WB). In one aspect, the D-dimer values ​​in oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are equivalent to those of conventionally stored blood. On the other hand, oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have D-dimer values ​​that are greater than the D-dimer value of conventionally stored blood.

[0091] The present disclosure provides and includes oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), whole blood (OR-WB) having coagulation parameters that are at least 75% of the coagulation parameters of conventionally stored whole blood as measured by a thrombin generation assay. In one aspect, the thrombin generation values ​​in oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are equivalent to those of conventionally stored blood. On the other hand, oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have thrombin generation values ​​that are greater than those of conventionally stored blood.

[0092] The present disclosure provides and includes oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), whose platelet function parameters are at least 75% of the platelet function parameters of conventional stored whole blood as measured by a platelet aggregometer. In one aspect, the platelet function parameters in oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are equivalent to those of conventionally stored blood. On the other hand, oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have platelet function parameters that are greater than those of conventionally stored blood.

[0093] The present disclosure provides and includes oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT), whose coagulation factor levels are at least 75% of the coagulation factor levels in traditionally stored blood. In one aspect, the coagulation factor levels are equivalent to traditionally stored blood. In other aspects, oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have platelet function parameters that are greater than the platelet function parameters of traditionally stored blood. Without being limited by theory, it is believed that oxidative degradation of coagulation factors can be prevented or reduced in oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), providing higher levels of coagulation factor activity. Methods for evaluating the effects of treatments on coagulability are known in the art, for example, Pidcoke et al., "Primary hemostatic capacity of whole blood: a comprehensive analysis of pathogen reduction and refrigeration effects over time," Transfusion 53: 137S-149S (2013).

[0094] The present disclosure provides and includes, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia and whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia and whole blood containing platelets (OCR-LRWB+PLT), the level of its factor V has a specific activity of at least 75% of the factor V activity level present in traditional stored blood. On the one hand, the specific activity of factor V is equivalent to traditional stored blood. In other aspects, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia and whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia and whole blood containing platelets (OCR-LRWB+PLT), the specific activity of its factor V is greater than the platelet function parameters of traditional stored blood. Methods for measuring the specific activity of factor V are known in the art.

[0095] The present disclosure provides and includes oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT), whose factor V level has a specific activity of at least 75% of the factor VIII activity level present in traditionally stored blood. In one aspect, the specific activity of factor VIII is comparable to traditionally stored blood. In other aspects, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia whole blood containing platelets (OCR-LRWB+PLT) have a specific activity of factor VIII that is greater than the platelet function parameters of traditionally stored blood. Methods for measuring factor VIII specific activity are known in the art. In one aspect, oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have a factor V specific activity of less than 40% after storage for 21 days.

[0096] The present disclosure provides and includes oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT), which have a certain level of antithrombin (AT) having a specific activity of at least 75% of the AT activity level in traditionally stored blood. In one aspect, the specific activity of AT is equivalent to that of traditionally stored blood. In other aspects, oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have a specific activity of AT greater than the platelet function parameters of traditionally stored blood. Methods for measuring specific AT activity are known in the art.

[0097] The present disclosure provides and includes oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) having a level of Factor XIV (autologous thrombin IIA or protein C) having a specific activity of at least 75% of the level of Factor XIV activity present in conventionally stored blood. In one aspect, the specific activity of Factor XIV is equivalent to that of conventionally stored blood. In other aspects, oxygen reduced leukopenic whole blood (OR-LRWB), oxygen reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have a specific activity of Factor XIV greater than platelet function parameters of traditionally stored blood. Methods for measuring the specific activity of Factor XIV are known in the art.

[0098] The present disclosure provides and includes oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT), which have a certain von Willebrand factor (vWF) level, and its specific activity is at least 75% of the vWF activity level in traditionally stored blood. In one aspect, the specific activity of vWF is equivalent to that of traditionally stored blood. In other aspects, oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB+PLT) have a specific activity of vWF greater than the platelet function parameters of traditionally stored blood. On the other hand, oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) have a specific activity of vWF that is less than the platelet function parameters of conventionally stored blood. Methods for measuring the specific activity of vWF are known in the art.

[0099] The present disclosure provides and includes methods for extending the shelf life of whole blood storage from the current 2 weeks to 3 weeks and beyond. The oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) of the present disclosure provide three-week patient results that are equivalent to patient results provided by whole blood that has been stored under conventional conditions for two weeks.

[0100] As provided herein, oxygen reduces the whole blood (OR-LRWB) of leukopenia, oxygen reduces the whole blood (OR-LRWB+PLT) of leukopenia and comprises platelet, oxygen and carbon dioxide reduce the whole blood (OCR-LRWB), oxygen and carbon dioxide reduce the whole blood (OCR-LRWB) of leukopenia and comprises platelet, compared with the blood of traditional storage, reduced the side effect of blood transfusion recipient.On the one hand, oxygen reduces the whole blood (OR-LRWB) of leukopenia, oxygen reduces the whole blood (OR-LRWB+PLT) of leukopenia and comprises platelet, oxygen and carbon dioxide reduce the whole blood (OCR-LRWB), oxygen and carbon dioxide reduce the whole blood (OCR-LRWB) of leukopenia and comprises platelet, compared with the blood of traditional storage, storage after two weeks inflammatory response reduces.In other respects, after three weeks, relative to the blood volume of traditional storage, inflammatory response is a reduction. On the one hand, oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) can be stored for more than three weeks and maintain the level of inflammatory response after two weeks compared to traditionally stored blood.

[0101] As provided herein, oxygen reduces the whole blood (OR-LRWB) of leukopenia, oxygen reduces the whole blood (OR-LRWB+PLT) of leukopenia and comprises platelet, oxygen and carbon dioxide reduce the whole blood (OCR-LRWB), oxygen and carbon dioxide reduce the whole blood (OCR-LRWB) of leukopenia and comprises platelet, compared with the blood of traditional storage, reduced the side effect of blood transfusion recipient.On the one hand, oxygen reduces the whole blood (OR-LRWB) of leukopenia, oxygen reduces the whole blood (OR-LRWB+PLT) of leukopenia and comprises platelet, oxygen and carbon dioxide reduce the whole blood (OCR-LRWB), oxygen and carbon dioxide reduce the whole blood (OCR-LRWB) of leukopenia and comprises platelet, compared with the blood of traditional storage, storage after two weeks immunomodulation reduces.In other respects, after three weeks, immunomodulation reduces relative to the blood of traditional storage. On the one hand, oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) can be stored for more than three weeks and maintain levels of immunomodulation compared to traditionally stored blood after two weeks.

[0102] The methods and whole blood products of the present invention provide improved patient outcomes when transfused. In particular, oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) provide improved survival of cancer patients provided in perioperative blood transfusion. In a certain aspect, oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) provide perioperative reduced mortality and improved survival for pancreatic cancer patients. Without being limited by theory, the reduced mortality is a result of reduced cytokine levels and improved oxygen transport and delivery due to increased 2,3-DPG and ATP levels.

[0103] In one aspect, the blood for transfusion into a cancer patient in need has reduced levels of activation-regulated cytokines, normal T-cell expressed and secreted (RANTES). In one aspect, the RANTES level is equivalent to the RANTES level at the start of storage. In another aspect, the RANTES level in oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen- and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), and oxygen- and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) is lower than the level of RANTES in conventionally stored blood. In one aspect, the level of RANTES is lower than the level of RANTES in conventionally stored blood during storage. In other aspects, RANTES does not increase during storage.

[0104] In one aspect, the blood transfused to a cancer patient in need thereof has reduced levels of CC chemokine, which is eotaxin, and in one aspect, the eotaxin that is reduced in oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood containing platelets (OR-LRWB+PLT), oxygen- and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), and oxygen- and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT) is eotaxin-1, also known as CC motif chemokine 11. In one aspect, the level of eotaxin is equivalent to the level of eotaxin present at the beginning of storage. On the other hand, the level of the eotaxin of oxygen-reduced leukopenic whole blood (OR-LRWB), oxygen-reduced leukopenic whole blood (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB+PLT) and the whole blood that comprises platelets is lower than the level of the eotaxin present in the blood of traditional storage.On the one hand, the level of eotaxin is lower than the level of the eotaxin present in traditional storage phase blood.In other aspects, eotaxin does not increase during storage.

[0105] The method and whole blood product of the present invention provide the multi-organ failure risk syndrome of reduction and improved patient outcome when input.Especially, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia and whole blood comprising platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia and whole blood comprising platelets (OCR-LRWB+PLT) provide the multi-organ failure risk syndrome of reduction in perioperative transfusion.In a certain aspect, oxygen reduced leukopenia whole blood (OR-LRWB), oxygen reduced leukopenia and whole blood comprising platelets (OR-LRWB+PLT), oxygen and carbon dioxide reduced leukopenia whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukopenia and whole blood comprising platelets (OCR-LRWB+PLT) provide the multi-organ failure risk syndrome of reduction in trauma patients during emergency treatment.

[0106] The present disclosure provides and includes a method of preparing oxygen-reduced leukopenic whole blood comprising obtaining a unit of whole blood comprising an anticoagulant, filtering the whole blood to produce leukopenic whole blood, depleting the leukopenic whole blood of oxygen, and storing the oxygen-reduced leukopenic whole blood under anaerobic conditions.

[0107] The present disclosure provides and includes a method for preparing oxygen-reduced leukopenic whole blood having a pre-stored oxygen saturation (SO2) of 30% or less. Whole blood obtained from a donor using venipuncture has an oxygen saturation ranging from about 30% to about 70% saturated oxygen (SO2). In certain aspects, the SO2 is reduced to 25% or less. In certain aspects, the SO2 is reduced to 20% or less. In certain aspects, sulfur dioxide is reduced to 15% or less. In other aspects, the SO2 is reduced to 10% or less. In other aspects, the SO2 is reduced to 5% or less.

[0108] Also provided and included in the present disclosure are compositions and methods for preparing oxygen-reduced and carbon dioxide-reduced leukopenic whole blood compositions. In certain aspects, the SO2 value is 20% or less and the partial pressure of carbon dioxide is less than 60 mmHg. In other aspects, the partial pressure of carbon dioxide is between 10 and 60 mmHg. In another aspect, the partial pressure of carbon dioxide is between 20 and 40 mmHg. Also included are whole blood compositions and methods that provide 15% or less SO2 and a partial pressure of carbon dioxide of 10-60 mmHg. In another aspect, the methods and compositions include whole blood products having 15% or less SO2 and a partial pressure of carbon dioxide of 20-40 mmHg. In another aspect, the blood compositions and methods of the present disclosure have 10% or less SO2 and a partial pressure of carbon dioxide of 10-60 mmHg. In other aspects, the blood compositions and methods of the present disclosure have 10% or less SO2 and a partial pressure of carbon dioxide of 20-40 mmHg. In yet another aspect, the blood compositions and methods of the present disclosure have 5% or less SO2 and a carbon dioxide partial pressure between 10 and 60 mmHg. In other aspects, the blood compositions and methods of the present disclosure have 5% or less SO2 and a carbon dioxide partial pressure between 20 and 40 mmHg.

[0109] Also provided and included in the present disclosure are compositions and methods for preparing oxygen-reduced and carbon dioxide-reduced leukopenic whole blood. In certain aspects, the SO2 value is 20% or less and the partial pressure of carbon dioxide is between 1 and 60 mmHg. In other aspects, the partial pressure of carbon dioxide is between 10 and 60 mmHg. In another aspect, the partial pressure of carbon dioxide is between 20 and 40 mmHg or 1 to 20 mmHg. Also included are whole blood compositions and methods that provide SO2 of 15% or less and a partial pressure of carbon dioxide of 10-60 mmHg. In certain aspects, the SO2 value is 15% or less and the partial pressure of carbon dioxide is between 1 and 60 mmHg. In another aspect, the methods and compositions include whole blood products having SO2 of 15% or less and a partial pressure of carbon dioxide of 20-40 mmHg or 1-20 mmHg. In another aspect, the blood compositions and methods of the present disclosure have SO2 of 10% or less and a partial pressure of carbon dioxide of 1 to 60 mmHg or 10 to 60 mmHg. In other aspects, the blood compositions and methods of the present disclosure have 10% or less SO2 and a carbon dioxide partial pressure of 20-40 mmHg or 1-20 mmHg. In other aspects, the blood compositions and methods of the present disclosure have 5% or less SO2 and a carbon dioxide partial pressure of 1 to 60 mmHg or 10 to 60 mmHg. In other aspects, the blood compositions and methods of the present disclosure have 5% or less SO2 and a carbon dioxide partial pressure of 20-40 mmHg or 1-20 mmHg.

[0110] It is worth noting that Figure 2A 、 2B As shown in Figures 3A and 3B, the ATP level in blood with reduced stored oxygen depends on the partial pressure of CO2. Specifically, depleting oxygen to about 10% SO2 and reducing carbon dioxide to about 25 mmHg resulted in an increase in 2,3-DPG levels that lasted for more than 21 days, while ATP decreased to about half of its initial value. Figure 2G and 3D Thus, the present disclosure provides and encompasses depleting oxygen to a SO2 level of about 5% and depleting carbon dioxide to a partial pressure of about 30 to 40 mmHg to produce 2,3-DPG in whole blood with increased levels of oxygen and reduced carbon dioxide, and maintaining at least 50% of the initial ATP concentration at day 20. In other aspects, the partial pressure of CO2 can be adjusted to maintain ATP levels at least 75% of the initial ATP value. Adjustment of CO2 levels can be determined experimentally by one of ordinary skill in the art, given the present disclosure.

[0111] Prolonged cryogenic storage under conventional conditions is known to impair the deformability of stored RBCs, potentially compromising their ability to perfuse the microvascular network and deliver oxygen to tissues and vital organs upon transfusion. Oxidative damage is believed to be a primary cause of the loss of RBC biomechanical function; therefore, storing RBCs under reduced oxygen (OR) and oxygen and carbon dioxide (OCR) conditions ameliorates oxidative damage, thereby better preserving native rheological properties than conventional (aerobic) storage. For this study, we utilized an in vitro microfluidic system that recapitulates an in vivo microvascular capillary bed to demonstrate the effects of reduced oxygen on stored cells.

[0112] The present disclosure provides and includes the method for managing blood bank, and this method improves the availability of blood products of trauma victims and the patient who needs multiple blood transfusions, and provides and preserves overall blood resources.This component blood product can be prepared from the whole blood preserved by the application, for blood transfusion or mixed with a large number of blood transfusion test kits.Except improved blood chemistry (hypohemolysis, improved 2,3-DPG etc.), this method also provides improved hemostasis and improved deformability.

[0113] In aspects according to the present specification, the method provides for maintaining an inventory of blood units comprising oxygen-reduced whole blood and an anticoagulant as described above, providing one or more blood units in the inventory for treating a patient, and recovering blood units from the inventory to prepare separated oxygen-reduced blood components, including oxygen-reduced plasma and oxygen-reduced leukopenia-containing packed red blood cells (OR-LRpRBC+PLT). In some aspects, the anticoagulant comprises citrate-phosphate-dextrose (CPD), citrate-phosphate-dextrose with adenine (CPDA-1), or CP2D.

[0114] In one aspect, the present disclosure provides a method for maintaining an inventory of blood units as described above, the blood units comprising oxygen- and carbon dioxide-reduced leukopenic whole blood and an anticoagulant, providing one or more blood units from the inventory to treat a patient, and recovering blood units from the inventory to prepare a component that separates oxygen- and carbon dioxide-reduced blood and oxygen- and carbon dioxide-reduced leukopenic packed red blood cells containing platelets (OCR-LRpRBC+PLT). In some aspects, the anticoagulant comprises citrate-phosphate-dextrose (CPD), citrate-phosphate-dextrose with adenine (CPDA-1), or the anticoagulant citrate phosphate diglucose (CP2D).

[0115] The specification further provides for the preparation of one or more massive transfusion kits as described below, which include oxygen-reduced plasma, oxygen-reduced leukopenia-containing packaged red blood cells (OR-LRpRBC+PLT), oxygen- and carbon dioxide-reduced plasma, and oxygen- and carbon dioxide-reduced leukopenia-containing packaged red blood cells (OCR-LRpRBC+PLT).

[0116] In aspects according to the present invention, unused blood in storage is recycled after a period of time. In certain aspects where the anticoagulant is CPD, blood units are recycled before three weeks of storage. In other aspects where the anticoagulant is CPDA-1, blood units are recycled before five weeks of storage. In other aspects, blood units are recycled after two weeks or less. In one aspect, blood unit recycling occurs between two days and one week. In another aspect, recycling occurs between two days and two weeks. In certain aspects, recycling occurs between one week and two weeks. The timing of recycling can vary depending on the blood facility's turnover and needs.

[0117] Although the recycling process is preferably performed under anaerobic conditions, the process can also be performed under aerobic conditions. Aerobic conditions may provide cost savings but may also be indicated in facilities with higher turnover. In high-turnover facilities, recovered blood components can be used shortly after the recycling process, and further storage of the blood under anaerobic conditions may provide little additional benefit.

[0118] The method for managing a blood bank further provides for the preparation of a large blood transfusion kit as described in detail below.

[0119] The present disclosure provides and includes: providing a method for supplying blood products for transfusion medicine, comprising depleting oxygen from leukopenic whole blood to prepare oxygen-reduced leukopenic whole blood (OR-LRWB+PLT), storing the oxygen-reduced leukopenic whole blood (OR-LRWB+PLT) for a period of time, and providing the stored blood to a patient in need. In certain aspects, the leukopenic whole blood step includes platelet reduction to produce the oxygen-reduced leukopenic whole blood (OR-LRWB).

[0120] The present disclosure provides and includes: providing a method for producing a blood product for transfusion medicine, comprising consuming oxygen and carbon dioxide from leukopenic whole blood to prepare oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB+PLT), storing the oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB+PLT) for a period of time, and providing the stored blood to a patient in need. In certain aspects, the leukopenic step includes platelet reduction to produce the oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB).

[0121] The present disclosure provides and includes: providing a method for supplying blood products for transfusion medicine, comprising depleting oxygen from leukopenic whole blood to prepare oxygen-reduced leukopenic whole blood (OR-LRWB+PLT), storing the oxygen-reduced leukopenic whole blood (OR-LRWB+PLT) for a period of time, and preparing oxygen-reduced leukopenic packaged red blood cells (OR-LRpRBC+PLT) containing platelets. In certain aspects, the leukopenic reduction step includes platelet reduction to produce oxygen-reduced leukopenic packaged red blood cells (OR-LRpRBC).

[0122] The present disclosure provides and includes: providing a method for producing a blood product for transfusion medicine, comprising consuming oxygen and carbon dioxide from leukocyte-reduced whole blood to prepare oxygen- and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB+PLT), storing the oxygen- and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB+PLT) for a period of time, and preparing oxygen- and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC+PLT) containing platelets. In certain aspects, the leukocyte reduction step includes platelet reduction to produce oxygen- and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC).

[0123] As provided herein, OR-LRpRBC+PLT, OR-LRpRBC, OCR-LRpRBC+PLT, and OCR-LRpRBC can be returned to the blood product supply store and stored for a period of time until the patient needs them. In aspects of the present disclosure, the total storage time as a whole blood product or as a packaged RBC product can be up to six weeks. In some aspects, the second storage period is between 2 and 4 weeks.

[0124] Methods for providing a blood product supply include providing oxygen or oxygen and carbon dioxide depletion. Methods for providing a blood product supply with oxygen levels are discussed in detail above. In certain aspects, the SO2 value is reduced to 20% or less, and the partial pressure of carbon dioxide is less than 60 mmHg. In other aspects, the partial pressure of carbon dioxide is between 10 and 60 mmHg. In another aspect, the partial pressure of carbon dioxide is between 20 and 40 mmHg. Also included are methods for providing an SO2 level of 15% or less and a partial pressure of carbon dioxide of 10-60 mmHg. In another aspect, the methods provide a blood product having an SO2 level of 15% or less and a partial pressure of carbon dioxide of 20-40 mmHg. In another aspect, the methods of the present disclosure provide a blood product having an SO2 level of 10% or less and a partial pressure of carbon dioxide of 10-60 mmHg. In other aspects, methods for providing a blood product supply provide an SO2 level of 10% or less and a partial pressure of carbon dioxide of 20-40 mmHg. In yet another aspect, the methods provide an SO2 level of 5% or less and a partial pressure of carbon dioxide of 10-60 mmHg. In other aspects, the method provides 5% or less SO2 and a carbon dioxide partial pressure of 20-40 mmHg.

[0125] The present disclosure provides and includes new blood components obtained in the blood component recovery process of OR-LRWB+PLT and OCR-LRWB+PLT. As mentioned above, although conventional whole blood products have an FDA-approved shelf life (3 weeks for WB in CPD and 5 weeks in CPDA1), clinicians using WB limit their shelf life to between 2 and 14 days. In conventional storage, blood is often discarded. In the present disclosure, OR-LRWB+PLT and OCR-LRWB+PLT can be processed using conventional component separation methods modified for keeping blood in an OR or OCR depleted state. Typically, the modification method is to combine a barrier layer impermeable to oxygen and oxygen and carbon dioxide to the component and combine features to prevent oxygen from entering. Suitable methods can be found in, for example, International Patent Application No. PCT / US2016 / 021794 filed on March 10, 2016 and International Patent Application No. PCT / US2016 / 029069 filed on April 22, 2016, both of which are incorporated herein by reference.

[0126] In accordance with an aspect of the present disclosure, a blood composition is provided comprising oxygen-reduced packaged red blood cells and a composition having a density of less than 1×10 5 In one aspect, the white blood cell level is less than 1×10 4 / L white blood cells. In aspects according to the present disclosure, the oxygen saturation of the oxygen reduced leukopenia packaged red blood cells containing platelets (OR-LRpRBC+PLT) is less than 30%. In one aspect, the oxygen saturation of the oxygen reduced leukopenia packaged red blood cells containing platelets (OR-LRpRBC+PLT) is less than 20%. In one aspect, the oxygen saturation of the oxygen reduced leukopenia packaged red blood cells containing platelets (OR-LRpRBC+PLT) is less than 10%. In another aspect, the oxygen saturation of the oxygen reduced leukopenia packaged red blood cells containing platelets (OR-LRpRBC+PLT) is less than 5%.

[0127] The present disclosure provides and includes packaged red blood cells (OCR-LRpRBC+PLT) that are oxygen and carbon dioxide reduced leukopenic and contain platelets with less than 30% sulfur dioxide and a carbon dioxide storage partial pressure of less than 60 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 30% and a carbon dioxide storage partial pressure of 20-40 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 30% and a carbon dioxide storage partial pressure between 0-20 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 20% and a carbon dioxide storage partial pressure of less than 60 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 20% and a carbon dioxide storage partial pressure between 20-40 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 20% and a carbon dioxide storage partial pressure between 0-20 mmHg. In another aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 15% and a carbon dioxide storage partial pressure of less than 60 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 15% and a carbon dioxide storage partial pressure of 20-40 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 15% and a carbon dioxide storage partial pressure between 0-20 mmHg. In another aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 10% and a carbon dioxide storage partial pressure between 60 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 10% and a carbon dioxide storage partial pressure between 20-40 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 10% and a carbon dioxide storage partial pressure between 0-20 mmHg. In another aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 5% and a carbon dioxide storage partial pressure of less than 60 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 5% and a carbon dioxide storage partial pressure of between 20-40 mmHg. In one aspect, the OCR-LRpRBC+PLT has an oxygen saturation of less than 5% and a carbon dioxide storage partial pressure of between 0-20 mmHg.

[0128] Oxygen-reduced leukopenia-containing packed red blood cells (OR-LRpRBC+PLT) and oxygen and carbon dioxide-reduced leukopenia-containing whole blood (OCR-LRWB+PLT) typically also contain an additive solution. Suitable additive solutions according to the present disclosure include AS-1, AS-3 ( ), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5, EAS61, OFAS1, OFAS3, and combinations thereof. In one aspect, the additive solution is added during component separation. In one aspect, the additive solution is AS-1. In another aspect, the additive solution is AS-3. In other aspects, the additive solution is SAGM.

[0129] The methods and compositions disclosed herein provide and include the preparation of a "massive transfusion kit" (MTK) having improved properties of a kit prepared from conventional components. The massive transfusion kit disclosed herein can be prepared in various configurations according to clinical needs. The MTK disclosed herein is stored under anaerobic or anaerobic and carbon dioxide-free conditions until ready for use. OR and OCR conditions can be maintained by sealing in an impermeable housing, with or without a suitable adsorbent material. The MTK disclosed herein can be reoxygenated before use, or used directly. In general, the specification stipulates that optimized massive transfusion kits can provide red blood cells with improved 2,3-DPG levels. Such kits are prepared from component blood products of oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB+PLT). Alternatively, a component blood product obtained from oxygen reduced leukopenic whole blood (OR-LRWB+PLT) containing platelets can be used to prepare a kit to generate a kit with a higher level of ATP. The kit prepared using the method of the present specification provides platelets suitable for hemostasis together with oxygen-reduced stored red blood cells. Therefore, the massive transfusion kit of the present specification can increase the availability of platelets without additional dilution, while further providing RBCs of higher quality (e.g., more deformable, more 2,3-DPG, less storage damage). Importantly, the recovery of blood components from the oxygen-reduced whole blood of the present disclosure increases the availability of transfusion products for trauma victims, saving and preserving valuable and limited resources. As described above, conventional massive transfusion kits include a certain volume of plasma, a certain volume of pRBC, and a certain volume of platelets in a 1:1:1 ratio, wherein the amounts of the three components correspond to a unit of "reconstituted blood" that is continuously or parallelly transfused to a patient in need. Reconstituted blood does not directly correspond to whole blood, does not contain additive solutions, and also has a higher anticoagulant content. Further reconstituted blood typically includes a larger volume than a typical unit of whole blood. The reconstituted blood of the present disclosure is improved over conventional reconstituted blood because it provides additional platelets in the pRBC fraction (e.g., oxygen and carbon dioxide reduced leukopenia packed red blood cells containing platelets (OCR-LRpRBC+PLT) and oxygen reduced leukopenia packed red blood cells containing platelets (OR-LRpRBC+PLT). Such cold-stored platelets are often referred to as platelet storage lesions (PSLs), and cold-stored platelets are rapidly expelled from the body's circulation. Importantly, cold-stored platelets have the ability to aggregate and are reported to have increased aggregation and resistance to decomposition. Therefore, blood components obtained from the oxygen-reduced whole blood of the present disclosure provide additional benefits during trauma transfusions, either alone or in combination with conventional platelets.

[0130] The present disclosure provides and includes a massive transfusion kit comprising a volume of oxygen-reduced leukopenic packed red blood cells containing platelets (OR-LRpRBC+PLT), oxygen- and carbon dioxide-reduced leukopenic whole blood containing platelets (OCR-LRWB+PLT), or a combination thereof. In one aspect, the massive transfusion kit provides a volume of plasma and a volume of LRpRBC+PLT. In one aspect, the volume of the plasma and the volume of LRpRBC+PLT is 1:1. In other aspects, the ratio of plasma to LRpRBC+PLT is between 1:1 and 1:2 by volume. In one aspect, the volume ratio of plasma to LRpRBC+PLT is approximately 1:2.

[0131] The present disclosure provides and encompasses massive transfusion kits comprising additional platelets with plasma and oxygen-reduced leukopenic packaged red blood cells (OR-LRpRBC+PLT) or oxygen- and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB+PLT).

[0132] The large-volume transfusion kit disclosed herein provides a certain volume of plasma. MTK's plasma can be fresh plasma or thawed fresh frozen plasma (FFP). The specification stipulates that MTK's plasma is obtained from a conventional source (e.g., non-oxygen-reduced) or an oxygen-reduced or oxygen and carbon dioxide-reduced source.

[0133] On the one hand, the plasma of the MTK of the present invention can be obtained from oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT). Without being limited by theory, plasma obtained from oxygen-reduced sources will have lower levels of storage damage, including, for example, lower levels of cytokines, isoprostanes, and microparticles. As provided herein, MTK with plasma, platelets, and pRBC according to the present specification is provided in a ratio of 1:1:1 or 1:1:2 by volume. It will be understood by those skilled in the art that the MTK of the present invention is designed to provide the equivalent of a blood unit, just like conventional MTK. It should be recognized that any arbitrary total volume can be selected while maintaining the necessary ratios mentioned to be equivalent to reconstituted blood.

[0134] Example

[0135] Example 1: Cytokine, cell-free hemoglobin, and isoprostane accumulation in packaged red blood cells during anaerobic storage

[0136] Fifteen pRBC units were collected from normal healthy donors and stored separately as described by Yoshida et al., "Anaerobic Storage of Red Blood Cells in a Novel Additive Solution Improves In vivo Recovery," Transfusion 49: 458-64 (2008). One unit was stored under standard blood bank conditions (control) and the other under anaerobic conditions (test). At 0, 1, 2, 3, and 6 weeks, samples were removed using a sterile connection device from the pRBC unit. Plasma samples were frozen for the following assays: 8-isoprostane F was analyzed by mass spectrometry using a Procarta immunoassay magnetic bead kit. 2α and single batch testing of 22 cytokines using cell-free hemoglobin by HemoCue plasma / photometer (HemoCue AB, Angelholm, Sweden).

[0137] like Figure 1A As shown, eotaxin reached statistically significant differences at week 2 (86.6 pg / ml-control (c), 64.9-test (t), p-value-0.00213, p < 0.05 was statistically significant; day 42 (292-c, 112-t; p = 0.000). As Figure 1B As shown, RANTES was different at all time points, starting from day 3 (374.6-c, 55.1-t), p-=0.00000; a very large difference was observed at day 42 (3371.6-c, 88.4-t; p<0.002). Figure 1C As shown, differences in cell-free hemoglobin were observed at week 2 (96.0 mg / d1-c, 41.7-t), p=0.00001; and day 42 (170-c, 63-t, p=0.0002). Figure 1D As shown, the difference in isoprostanol was shown on the 3rd day of storage (45.5 pg / ml-c, 32.1-t), p=0.00689; and on the 42nd day (101.9-c, 64.7-t, p=0.0048).

[0138] Example 2: Whole Blood Collection, Whitening and Gas Consumption

[0139] Blood units from donor patients were collected according to standard protocols into an anticoagulant solution containing CPDA1 or CPDA, including collection heparin tubing. Collected anticoagulant blood was leukopenic according to the manufacturer's instructions within 4 hours of the initial blood draw. Baseline ABL90 blood gas and metabolic parameters were determined from donor heparin tubing and whole blood products according to standard procedures. Please refer to the BSL Handbook Procedure BSL-P024: Procedure Manual and RadiometerABL90FLEX Gas Analyzer instructions.

[0140] Anaerobic controls were prepared from each leukoreduced blood by transferring 120 ml of LRWB / CPDA-1 or LRWB / CPD into a 150 mL transfer bag, labeled as appropriate, and placed at room temperature (15-30°C).

[0141] The remaining LRWB / CPDA-1 or LRWB / CPD was processed for oxygen or oxygen and carbon dioxide depletion by transferring to a blood processing bag connected to a Sorin D100 and processed at a flow rate of 700 ml / min for 5 minutes without gas generation (BOF-processed control). 120 g of the resulting BOF-processed blood was transferred to a 300 ml transfer bag that had been stored under anaerobic conditions and labeled as a BOF-processed control. The remaining LRWB / CPDA-1 or LRWB / CPD was processed on the Sorin D100 at a peak flow rate of 700 ml / min with a gas flow rate of 3 liters / min and a gas composition of 5% CO2 and 95% N2 until the blood reached ~5% SO2. Blood gas values ​​were measured at 3 to 5 minute intervals on a Radiometer ABL90FLEX gas analyzer. To reduce carbon dioxide levels, the gas mixture was switched to 100% N₂ for 1-4 minutes until SO₂ reached 5±1% and pCO₂ reached 30±3 mmHg. Blood gas values ​​were monitored every 15-30 seconds to monitor the rate of deoxygenation. 120 g of the resulting oxygenated and carbon dioxide-reduced LRWB / CPDA-1 or LRWB / CPD was transferred to a 300 ml transfer bag previously stored under anaerobic conditions as described above and labeled ("C"). Further processing of the LRWB / CPDA-1 or LRWB / CPD was performed on a Sorin D100 using 99% N₂ and 1% O₂ at a flow rate of 700 ml / min until the LRWB / CPDA-1 or LRWB / CPD reached 5±1% SO₂ and pCO₂ reached 7±3 mmHg. Transfer 120 g of the resulting oxygen- and carbon dioxide-reduced LRWB / CPDA-1 or LRWB / CPD to a 300 ml transport bag previously stored under anaerobic conditions as described above and labeled ("D"). Additional samples were processed as described above using a new Sorin D100. Immediately following the preparation of each sample, ABL90 blood gas levels were determined according to the manufacturer's instructions to establish baseline SO2 and pCO2 levels (e.g., T0). See BSL Handbook Procedures. Samples for cytokine analysis were collected and stored at -80°C for later analysis.

[0142] All samples were analyzed as provided in Example 6 below.

[0143] Example 3: Storage of Anaerobic Test Products

[0144] Oxygen-reduced and oxygen- and carbon dioxide-reduced blood transfer bags were wrapped in mesh, secured with elastic, and placed in anaerobic canisters along with four sorbent bags (Mitsubishi, SS-300). The canisters were sealed and purged of air using an Alicat gas handling system. See BSL Handbook Procedure BSL-P040: Procedure for Placing Blood Products in Anaerobic Storage in Canisters. Anaerobic and aerobic blood was placed in a blood bank refrigerator at 1 to 6°C. The canister pressure gauge was monitored daily to ensure a reading of 5 ± 1 psi. Canisters that fell below 2 psi were adjusted to standard procedures. See BSL Handbook Procedure BSL-P040: Procedure for Placing Blood Products in Anaerobic Storage in Canisters.

[0145] Example 4: Example Test

[0146] Samples were tested at designated time points: day 0 (T0) post-treatment, day 1, week 1, week 2, and week 3. For a given test, samples could be tested fresh or frozen. Tests included complete blood count (CBC) and thromboelastography (TEG).

[0147] Immediately prepare platelet-rich plasma (PRP) for platelet aggregation according to the manufacturer's instructions.

[0148] Coagulation screening and additional testing were performed according to the manufacturer's instructions.

[0149] Immediately prepare cytokine samples according to the manufacturer's instructions.

[0150] Example 5: ATP sampling and measurement

[0151] ATP measurement samples are processed by deproteinization and precipitation. 1 ml of sample (e.g., LRWB / CPD or LRWB / CPDA-1 or the above samples) is precipitated with 1.0 ml of ice-cold trichloroacetic acid (TCA) (12% w / v) and vortexed for 15 to 30 seconds and incubated on ice for 5 minutes. The tube containing the TCA / sample mixture is centrifuged at 3600 g for 5 minutes at 4°C. The sample is processed immediately to minimize exposure to TCA. The clarified supernatant is transferred to a pre-cooled tube and quickly frozen in a dry ice alcohol bath and stored at -70°C.

[0152] Example 6: Improved deformability in RBCs stored under reduced oxygen conditions

[0153] Nine (9) individual units of whole blood were obtained from healthy consenting volunteers via a standard 500 mL blood donation. The whole blood donation was processed into leukoreduced red blood cell (LR-RBC) units according to standard AABB / FDA guidelines; the resulting units were then split in half. One half of the units was O2 and CO2 reduced as described in Examples 2 and 3.

[0154] The obtained samples were placed in anaerobic low temperature storage, while the latter half was placed in conventional aerobic low temperature storage. Paired red blood cell units were stored in a blood bank refrigerator and evaluated weekly throughout the 6-week storage period. Before testing, the hematocrit of all RBC samples was adjusted to 40% using normal saline (0.9% NaCl; RBC-S). The deformability of RBC-S at the beginning of the study and during the study was determined as described in International Patent Publication No. WO2013 / 177339 published on November 28, 2013. High-speed image sequences (~150FPS) of blood samples passing through an artificial microvascular network (AMVN) chip were recorded. The amount of time it takes for non-deformable cells to pass through the network and the frequency of blood flooding through the network (blocking frequency) were determined.

[0155] For both oxygenated and carbon dioxide-controlled blood, the overall bulk perfusion rate through the AMVN system was consistently higher compared to the aerobically stored units, while the total occlusion time for oxygen-reduced RBCs was consistently lower (Table 1). These results suggest that the reduction in oxygen content in LR-RBC units mitigates the deterioration of RBC biomechanical properties during cryogenic storage.

[0156] The hypoxia and storage process significantly reduced the rate at which the rheological properties of RBCs deteriorated during low-temperature storage and was able to maintain more physiologically relevant biomechanical properties of RBCs during storage. Combined with the benefits of whole blood transfusion, improved RBC deformability suggests that preserved RBC function will improve RBC retention after transfusion and increase the ability of transfused RBCs to diffuse through the microvasculature.

[0157] Table 1: Perfusion Rates of Blood Cells After Oxygen-Reduced Storage

[0158]

[0159]

[0160] Example 7: Deoxygenation of platelets does not hinder hemostatic properties

[0161] Eight (8) units of whole blood (WB) were obtained from healthy consenting volunteers through standard 500 mL blood donations. The donated whole blood was collected in CPDA-1 anticoagulant and filtered through a platelet-retaining filter ( WB-SP) (LRWB; Terumo Medical Corporation). The resulting filtered units were then divided in half. One half of the units was placed in conventional, aerobic, low-temperature storage, while the other half was further divided for anaerobic, low-temperature storage. The anaerobic storage units were either oxygen-reduced (OR-LRWB) or oxygen and carbon dioxide-reduced (OCR-LRWB). The anaerobically stored units were processed using a Sorin D100 membrane oxygenator to produce anaerobic units with approximately 5% SO2 and approximately 35 mm Hgp CO2. The resulting anaerobic units were placed in standard PVC bags and stored in anaerobic tanks containing oxygen sorbent and nitrogen. Paired leukocyte-reduced platelet units were stored every 21 days as described below.

[0162] Metabolic parameters including percentage of hemolysis (PlasmaLow, Angelholm, Sweden), ATP (DiaSys, Flacht, Germany), and 2,3-DPG (Sigma-Aldrich, St. Louis, MO) were assessed according to the manufacturer's instructions. Figure 4A As shown, the reduced ATP level in stored OCR-LRWB remained unchanged, but the ATP level in stored OR-LRWB increased compared to conventionally stored LRWB (solid line). Figure 4B As shown in Figure 2, the stored OCR-LRWB and OR-LRWB maintained the 2,3-DPG level for up to 21 days compared to the conventionally stored LRWB. Figure 4C As shown, there was no significant change in blood stability when storing OR-LRWB and storing OCR-LRWB were compared with conventionally stored LRWB (solid line).

[0163] Plasma coagulation parameters were evaluated in conventionally stored LRWB and OCR-LRWB by assessing prothrombin time (PT), activated partial prothrombin time (aPTT), and fibrinogen and D-dimer levels. As shown in Figure 5, aPTT and PT were slightly, but not significantly, prolonged in conventionally stored LRWB (solid lines). In addition, no evidence of coagulation activation was observed, as evidenced by similar fibrinogen and D-dimer levels.

[0164] Plasma coagulation factors of conventionally stored LRWB and OCR-LRWB were further evaluated by determining the activity levels of factors V, VIII, protein C activity, protein S activity, and von Willebrand factor (vWF). Protein C and protein S assays were performed using ACL according to the manufacturer's instructions. (Instrumentation Laboratory) and (DiagnosticaStago, Inc.) As shown in Figure 6, the levels of factor V, factor VIII, protein C activity, protein S activity, and vWF did not change significantly in anaerobic, low-temperature stored OCR-LRWB (dashed line) compared with conventionally stored WB (solid line).

[0165] Conventional stored LRWB and OCR-LRWB were analyzed using a Haemoscope according to the manufacturer's instructions. The coagulation function was further evaluated by Haemonetics analyzer and thromboelastography (TEG). Figures 7A to 7D As shown, no significant differences were observed in propagation (TEG angle), amplification (TEGK), maximum amplitude (TEGMA), or reaction time (TEG R) in OCR-LRWB (dashed line) compared with conventionally stored LRWB (solid line).

[0166] Although the present disclosure has been described with reference to specific embodiments, it will be understood by those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof without departing from the scope of the present disclosure. Additionally, many modifications may be made to adapt a particular situation or material to the teachings of the present disclosure without departing from the scope of the present disclosure.

[0167] Therefore, it is intended that the disclosure not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this disclosure, but that the disclosure will include all embodiments falling within the scope and spirit of the appended claims.

[0168] This application also relates to the following embodiments:

[0169] 1. A method for improving the survival of a patient requiring multiple transfusions comprising providing oxygen-reduced stored red blood cells (OR-stored RBCs) to a patient requiring a medical procedure.

[0170] 2. The method of embodiment 1, wherein the OR-stored RBCs are oxygen and carbon dioxide reduced (OCR-stored RBCs).

[0171] 3. The method of embodiment 1 or 2, wherein the OR-stored RBCs comprise reduced levels of unbound factor RANTES compared to conventionally stored red blood cells (stored RBCs).

[0172] 4. The method of any one of embodiments 1 to 3, wherein the level of RANTES is less than 500 pg / ml after 21 days.

[0173] 5. The method of any one of embodiments 1 to 4, wherein the level of RANTES is less than 300 pg / ml after 21 days of storage under deoxygenated conditions.

[0174] 6. The method of any one of embodiments 1 to 5, wherein the OR-stored RBCs comprise reduced levels of eotaxin compared to conventionally stored red blood cells (stored RBCs).

[0175] 7. The method of any one of embodiments 1 to 6, wherein the level of eotaxin is less than 150 pg / ml after 21 days of storage under deoxygenated conditions.

[0176] 8. The method of any one of embodiments 1 to 7, wherein the level of eotaxin is less than 100 pg / ml after 21 days of storage under deoxygenated conditions.

[0177] 9. The method of any one of embodiments 1 to 8, wherein the OR-stored RBCs are selected from the group consisting of oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen-reduced leukocyte-reduced packaged red blood cells (OR-LRpRBC), oxygen-reduced leukocyte-reduced packaged red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), oxygen and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), oxygen and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC+PLT), and combinations thereof.

[0178] 1 0. The method described in any one of embodiments 1 to 9, wherein the OR-stored RBCs include oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT).

[0179] 11. The method of any one of embodiments 1 to 10, wherein the patient requiring multiple transfusions is a trauma patient, a transplant patient, a cardiac surgery patient, an obstetric patient, a GI surgery patient, or an orthopedic surgery patient.

[0180] 12. The method of any one of embodiments 1 to 11, wherein the trauma patient is a hemorrhagic trauma patient or a blunt trauma patient.

[0181] 13. The method of any one of embodiments 1 to 12, wherein the patient is a cancer patient and the OR-stored RBCs are oxygen-reduced leukopenia packaged red blood cells (OR-LRpRBC), oxygen-reduced leukopenia packaged red blood cells containing platelets (OR-LRpRBC+PLT), oxygen- and carbon dioxide-reduced leukopenia packaged red blood cells (OCR-LRpRBC), or oxygen- and carbon dioxide-reduced leukopenia packaged red blood cells containing platelets (OCR-LRpRBC+PLT).

[0182] 14. A method for improving the survival of cancer patients requiring perioperative transfusions comprising providing oxygen-reduced stored red blood cells (OR-stored RBCs) to a patient requiring a medical procedure.

[0183] 15. The method of embodiment 14, wherein the OR-stored RBCs are oxygen- and carbon dioxide-reduced red blood cells (OCR-stored RBCs).

[0184] 16. The method of embodiment 14 or 15, wherein the OR-stored RBCs are oxygen-reduced leukopenia packaged red blood cells (OR-LRpRBC), oxygen-reduced leukopenia packaged red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide-reduced leukopenia packaged red blood cells (OCR-LRpRBC), or oxygen and carbon dioxide-reduced leukopenia packaged red blood cells containing platelets (OCR-LRpRBC+PLT).

[0185] 17. The method of any one of embodiments 14 to 16, wherein the OR-stored RBCs comprise reduced levels of the factor RANTES or eotaxin compared to non-oxygen-reduced stored red blood cells (stored RBCs).

[0186] 18. The method of any one of embodiments 14 to 17, wherein the level of RANTES is less than 500 pg / ml after 21 days, or the level of eotaxin is less than 150 pg / ml after 21 days of storage under deoxygenated conditions.

[0187] 19. A method for reducing cytokines in stored blood, comprising:

[0188] Blood was collected into an anticoagulant solution;

[0189] reducing leukocytes from the collected blood;

[0190] Reducing pre-storage oxygen saturation (SO2) to 30% or less; and

[0191] The collected oxygen-reduced blood is stored under anaerobic conditions to prepare oxygen-reduced stored blood.

[0192] 20. The method of embodiment 19, further comprising removing the plasma and adding additional solution to prepare oxygen-reduced packed red blood cells containing platelets (OR-pRBC+PLT).

[0193] 21. The method of embodiment 19, wherein the reduction of the white blood cells further comprises reducing platelets.

[0194] 22. The method of embodiment 21, further comprising removing the plasma and adding additional solution to prepare oxygen-reduced packaged red blood cells (OR-pRBC).

[0195] 23. The method of any one of embodiments 19 to 22, wherein the level of one or more cytokines is reduced compared to non-oxygen-reduced stored blood cells (stored RBCs).

[0196] 24. The method of any one of embodiments 19 to 23, wherein the cytokine is selected from the group consisting of: monocyte chemoattractant protein-1 (MCP-1), regulated chemokine expressed and secreted on activated normal T cells (RANTES), angiogenin, tumor necrosis factor-α (TNF-α), epidermal growth factor (EGF), soluble CD40 ligand (sCD40L) and platelet-derived growth factor (PDGF).

[0197] 25. The method of embodiment 19, wherein reducing the pre-storage oxygen saturation further comprises reducing the pre-storage carbon dioxide partial pressure to less than 60 mmHg.

[0198] 26. The method of embodiment 25, further comprising removing the plasma and adding additional solution to produce oxygen- and carbon dioxide-reduced packed red blood cells containing platelets (OCR-pRBC+PLT).

[0199] 27. The method of embodiment 25, wherein the reduction of the white blood cells further comprises reducing platelets.

[0200] 28. The method of embodiment 27, further comprising removing the plasma and adding additional solution to prepare oxygen-reduced packaged red blood cells (OR-pRBC).

[0201] 29. The method of any one of embodiments 19 or 25 to 28, wherein the oxygen-reduced stored blood comprises a reduced level of factor RANTES compared to non-oxygen-reduced stored red blood cells (stored red blood cells).

[0202] 30. The method of any one of embodiments 19 or 25 to 29, wherein the oxygen-reduced stored blood comprises a reduced level of eotaxin compared to non-oxygen-reduced stored red blood cells (stored red blood cells).

[0203] 31. The method of embodiment 19, wherein the oxygen-reduced stored blood has an elevated level of 2,3-DPG, wherein the oxygen-reduced leukopenic blood has a 2,3-DPG level that is higher than the initial 2,3-DPG level in the leukopenic blood over 15 days.

[0204] 32. The method of embodiment 19 or 31, wherein the 2,3-DPG level is at least 80% or more higher than the 2,3-DPG level in the blood at day 0.

[0205] 33. The method of any one of embodiments 19, 31 to 32, wherein the 2,3-DPG level on day 21 is between 5 and 20 μmol 2,3-DPG / gHb.

[0206] 34. The method of any one of embodiments 19 to 33, wherein the oxygen-reduced stored blood is selected from the group consisting of oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen-reduced leukocyte-reduced packed red blood cells (OR-LRpRBC), oxygen-reduced leukocyte-reduced packed red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced packed red blood cells (OCR-LRpRBC), oxygen and carbon dioxide-reduced leukocyte-reduced packed red blood cells (OCR-LRpRBC), oxygen and carbon dioxide-reduced leukocyte-reduced packed red blood cells (OCR-LRpRBC+PLT), and combinations thereof.

[0207] 35. A blood composition for transfusion into a trauma patient in need thereof, comprising oxygen-reduced leukopenic whole blood (OR-LRWB) in an anticoagulant solution having a pre-storage and storage oxygen saturation (SO2) of 30% or less, which has a 2,3-DPG level over 15 days that is higher than the initial 2,3-DPG level in said oxygen-reduced leukopenic whole blood, or oxygen- and carbon dioxide-reduced leukopenic whole blood (OCR-LRWB) having a pre-storage and storage oxygen saturation (SO2) of 30% or less and a pre-storage and storage carbon dioxide partial pressure of less than 60 mmHg, wherein said OCR-LRWB has an adenosine triphosphate (ATP) level of at least 3 μmol / gHb.

[0208] 36. The blood composition of embodiment 35, wherein the 2,3-DPG level in the OR-LRWB is at least 80% or more higher than the 2,3-DPG level in the blood at day 0.

[0209] 37. The blood composition of embodiment 35 or 36, wherein the 2,3-DPG level of the OR-LRWB is at least 5 to 20 DPG μmol / gHb.

[0210] 38. The blood composition of any one of embodiments 35 to 37, wherein the pre-storage oxygen saturation is less than 20%, less than 10%, or less than 5%.

[0211] 39. The blood composition of any one of embodiments 35 to 38, wherein the pre-storage partial pressure is between 1 and 60 mmHg, 10 and 60 mmHg, 20 and 40 mmHg, 1 and 20 mmHg.

[0212] 40. The blood composition of any one of embodiments 35 to 39, wherein the anticoagulant solution is citrate-phosphate-dextrose containing adenine (CPDA1) or citrate-phosphate-dextrose (CPD).

[0213] 41. The blood composition of any one of embodiments 35 to 40, wherein the oxygen reduced leukocyte reduced whole blood (OR-LRWB) or oxygen and carbon dioxide reduced leukocyte reduced whole blood (OCR-LRWB) has reduced levels of biological response modifiers (BRMs) selected from the group consisting of: cytokines, chemokines, isoprostanes, and oxidized lipid products.

[0214] 42. The blood composition of embodiment 41, wherein a patient transfused with the oxygen reduced leukocyte reduced whole blood (OR-LRWB) or oxygen and carbon dioxide reduced leukocyte reduced whole blood (OCR-LRWB) has a reduced inflammatory response.

[0215] 43. The blood composition of embodiment 41 or 42, wherein a patient transfused with the oxygen reduced leukopenic whole blood (OR-LRWB) or oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB) has reduced immunomodulation.

[0216] 44. The blood composition of any one of embodiments 41 to 43, wherein a patient transfused with the oxygen reduced leukopenic whole blood (OR-LRWB) or oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB) has a reduced risk of multiple organ failure.

[0217] 45. The blood composition of any one of embodiments 41 to 44, wherein a patient transfused with the oxygen reduced leukopenic whole blood (OR-LRWB) or oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB) has a reduced risk of sepsis.

[0218] 46. ​​The blood composition of any one of embodiments 41 to 45, wherein a patient transfused with the oxygen reduced leukopenic whole blood (OR-LRWB) or oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB) has a reduced risk of infection.

[0219] 47. The blood composition of any one of embodiments 41 to 46, wherein a patient transfused with the oxygen reduced leukopenic whole blood (OR-LRWB) or oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB) has a reduced risk of death.

[0220] 48. The blood composition of any one of embodiments 41 to 47, wherein a patient transfused with the oxygen reduced leukopenic whole blood (OR-LRWB) or oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB) has increased red blood cell (RBC) deformability.

[0221] 49. The blood composition of any one of embodiments 41 to 48, wherein the blood has equivalent or better coagulation parameters as measured by thromboelastography (TEG) compared to non-oxygen reduced leukocyte reduced whole blood (OR-LRWB) or non-oxygen and carbon dioxide reduced leukocyte reduced whole blood (OCR-LRWB).

[0222] 50. The blood composition of any one of embodiments 41 to 49, wherein the blood has equivalent or better coagulation parameters as compared to non-oxygen reduced leukocyte reduced whole blood (OR-LRWB) or non-oxygen and carbon dioxide reduced leukocyte reduced whole blood (OCR-LRWB), as determined by PT / PTT.

[0223] 51. The blood composition of any one of embodiments 41 to 50, wherein the blood has equivalent or better coagulation parameters as measured by a platelet aggregometer compared to non-oxygen reduced leukocyte-reduced whole blood (OR-LRWB) or non-oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB).

[0224] 52. The blood composition of any one of embodiments 41 to 51, wherein the blood has equivalent or better levels of clotting factors, including Factor V, Factor VIII, AT, Protein C, or vWF, compared to non-oxygen reduced leukopenic whole blood (OR-LRWB) or non-oxygen and carbon dioxide reduced leukopenic whole blood (OCR-LRWB).

[0225] 53. The blood composition of any one of embodiments 41 to 52, wherein the blood is safe for transfusion into a patient in need thereof for at least 3 weeks.

[0226] 54. A method of reducing an inflammatory response in a patient receiving a blood transfusion, comprising transfusing a reduced oxygen blood product to a patient in need thereof, wherein the reduced oxygen blood product has reduced levels of inflammatory factors after storage under anaerobic conditions.

[0227] 55. The method of embodiment 54, wherein the reduced level of an inflammatory factor is reduced levels of eotaxin or RANTES compared to non-oxygen-reduced stored red blood cells.

[0228] 56. The method of embodiment 54 or 55, wherein the blood product is selected from the group consisting of: oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen reduced leukocyte-reduced packaged red blood cells (OR-LRpRBC), oxygen reduced leukocyte-reduced packaged red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), oxygen and carbon dioxide reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), oxygen and carbon dioxide reduced leukocyte-reduced packaged red blood cells containing platelets (OCR-LRpRBC+PLT), plasma, and combinations thereof.

[0229] 57. A method of reducing immunomodulation in a patient receiving a blood transfusion, comprising transfusing an oxygen-reduced blood product to a patient in need thereof, wherein said oxygen-reduced blood product has reduced levels of cytokines after storage under anaerobic conditions, wherein said levels of said cytokines are compared to a non-oxygen-reduced blood product.

[0230] 58. The method of embodiment 57, wherein the cytokine is selected from the group consisting of: monocyte chemoattractant protein-1 (MCP-1), regulated chemokine expressed and secreted on activated normal T cells (RANTES), angiogenin, tumor necrosis factor-α (TNF-α), epidermal growth factor (EGF), soluble CD40 ligand (sCD40L) and platelet-derived growth factor (PDGF).

[0231] 59. The method of embodiment 57 or 58, wherein the cytokine is eotaxin or RANTES.

[0232] 60. The method described in any one of embodiments 57 to 59, wherein the blood product is selected from the group consisting of oxygen reduced leukocyte-reduced whole blood (OR-LRWB), oxygen reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen reduced leukocyte-reduced packaged red blood cells (OR-LRpRBC), oxygen reduced leukocyte-reduced packaged red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), oxygen and carbon dioxide reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), oxygen and carbon dioxide reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), oxygen and carbon dioxide reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC+PLT), plasma, platelet concentrates, and combinations thereof.

[0233] 61. A method for improving oxygen perfusion in a patient in need thereof, comprising transfusing a reduced oxygen blood product to a patient in need thereof, wherein the reduced oxygen blood product has a higher RBC deformation than a conventionally stored blood product.

[0234] 62. The method of embodiment 61, wherein the oxygen-reduced blood product is selected from the group consisting of oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen-reduced leukocyte-reduced packaged red blood cells (OR-LRpRBC), oxygen-reduced leukocyte-reduced packaged red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), oxygen and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), oxygen and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC+PLT), plasma, platelet concentrates, and combinations thereof.

[0235] 63. The method of embodiment 61 or 62, wherein the oxygen-reduced blood product is a recovered blood product comprising oxygen-reduced leukocyte-reduced packaged red blood cells (OR-LRpRBC), oxygen-reduced leukocyte-reduced packaged red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced packaged red blood cells (OCR-LRpRBC), or oxygen and carbon dioxide-reduced (OCR-LRpRBC+PLT), obtained by fractionating, plasma-removing, and adding additional solutions from oxygen-reduced leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB), and oxygen and carbon dioxide-reduced leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) after storage for at least one week.

[0236] 64. The method of any one of embodiments 61 to 63, wherein the recovered blood product is stored for up to 42 days, 56 days, or 64 days.

[0237] 65. Methods of managing a blood bank, including:

[0238] Maintaining an inventory of blood units containing oxygen-reduced whole blood and anticoagulant, or oxygen-reduced leukopenic whole blood and anticoagulant;

[0239] providing one or more of said blood units from said inventory for use in treating a patient;

[0240] Blood units are recovered from the bank to prepare componentized oxygen-reduced blood units.

[0241] 66. The method of embodiment 65, wherein the component-separated oxygen-reduced blood unit comprises oxygen-reduced leukopenic packed red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide-reduced leukopenic packed red blood cells containing platelets (OCR-LRpRBC+PLT), oxygen-reduced plasma, or a combination thereof.

[0242] 67. The method of embodiment 65 or 66, wherein the component-separated oxygen-reduced blood units are recovered to prepare a large blood transfusion kit.

[0243] 68. The method of any one of embodiments 65 to 67, wherein the anticoagulant comprises citrate-phosphate-dextrose (CPD) or citrate-phosphate-dextrose containing adenine (CPDA-1).

[0244] 69. The method of any one of embodiments 65 to 68, wherein said salvaging of the blood unit is performed 3 weeks prior to storage for a blood unit containing CPD and 5 weeks prior to storage for a blood unit containing CPDA1.

[0245] 70. The method of any one of embodiments 65 to 69, wherein said salvaging of blood units is performed at 2 weeks.

[0246] 71. The method of any one of embodiments 65 to 70, wherein said recovering a blood unit is performed under anaerobic conditions.

[0247] 72. The method of any one of embodiments 65 to 71, wherein the componentized oxygen-reduced blood unit is stored for up to 6 weeks after adding additional solution.

[0248] 73. The method of any one of embodiments 65 to 72, further comprising preparing a massive transfusion kit comprising a volume of plasma and leukopenia- and platelet-containing packaged red blood cells (LRpRBC+PLT).

[0249] 74. The method of embodiment 73, wherein the ratio of plasma to LRpRBC+PLT is 1:1 by volume.

[0250] 75. The method of embodiment 73 or 74, wherein the ratio of plasma to LRpRBC+PLT is 1:2 by volume.

[0251] 76. The method of embodiment 73, wherein the massive blood transfusion kit further comprises a certain volume of platelets, wherein the ratio of the plasma, the platelets and the LRpRBC+PLT is 1:1:1 by volume.

[0252] 77. The method of embodiment 73 or 76, wherein the massive blood transfusion kit further comprises a certain volume of platelets, wherein the ratio of the plasma, the platelets and the LRpRBC+PLT is 1:1:2 by volume.

[0253] 78. A method of providing a supply of blood products for use in transfusion medicine, comprising:

[0254] Depleting whole blood of oxygen or carbon dioxide to prepare oxygen-reduced whole blood (or oxygen- and carbon dioxide-reduced whole blood); and

[0255] storing the oxygen or oxygen and carbon dioxide reduced whole blood for a period of time and providing the stored blood to a patient in need thereof; or

[0256] The oxygenated or oxygen and carbon dioxide reduced whole blood is stored for a period of time, and oxygenated or oxygen and carbon dioxide reduced packaged red blood cells are prepared.

[0257] 79. The method of embodiment 78, wherein the oxygen and carbon dioxide reduced packaged red blood cells are stored for a second period of time and then provided to a patient in need thereof.

[0258] 80. The method of embodiment 78 or 79, wherein the second period of time is up to 6 weeks.

[0259] 81. A blood composition comprising oxygen-reduced packed red blood cells and platelets containing less than 1×10 5 / L white blood cells.

[0260] 82. The blood composition of embodiment 81, wherein the composition comprises less than 1×10 4 / L white blood cells.

[0261] 83. The blood composition of embodiment 81 or 82, further comprising an additional solution selected from the group consisting of: AS-1, AS-3 ( ), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5, EAS61, OFAS1, OFAS3 and their combinations.

[0262] 84. A massive blood transfusion kit comprising a certain volume of plasma and a certain volume of LRpRBC+PLT.

[0263] 85. The massive blood transfusion kit of embodiment 84, wherein the ratio of said plasma to said LRpRBC+PLT is 1:1 by volume.

[0264] 86. The massive blood transfusion kit of embodiment 84 or 85, wherein the ratio of said plasma to said LRpRBC+PLT is 1:2 by volume.

[0265] 87. The massive blood transfusion kit of embodiment 84 further comprises a volume of platelets, wherein the ratio of the plasma, the platelets and the LRpRBC+PLTs is 1:1:1 by volume.

[0266] 88. The massive blood transfusion kit of embodiment 84 or 87 further comprises a volume of platelets, wherein the ratio of the plasma, the platelets, and the LRpRBC+PLTs is 1:1:1 by volume.

Claims

1. A method for reducing the level of one or more cytokines in stored whole blood, comprising: Whole blood was collected into anticoagulant solution; reducing leukocytes from the collected whole blood to produce leukoreduced platelet-containing whole blood (LRWB+PLT); reducing the pre-storage oxygen saturation (SO2) to 30% or less to produce an oxygen-reduced LRWB+PLT (OR-LRWB+PLT); and storing the OR-LRWB+PLT under anaerobic conditions to prepare oxygen-reduced leukopenic whole blood containing platelets, wherein the level of cytokines in the stored OR-LRWB+PLT is reduced compared to conventionally stored non-oxygen-reduced leukopenic whole blood containing platelets (non-OR-LRWB+PLT), wherein the one or more cytokines are regulated chemokine expressed and secreted on activated normal T cells (RANTES), eotaxin, or both, and wherein the levels of the one or more cytokines in the stored OR-LRWB+PLTs are reduced compared to the levels of the same one or more cytokines in conventionally stored non-OR-LRWB+PLTs.

2. The method of claim 1, further comprising removing plasma and adding an additive solution to the stored OR-LRWB+PLT to prepare oxygen-reduced leukopenic stored packed red blood cells containing platelets (OR-LRpRBC+PLT).

3. The method of claim 1, wherein said reduction of said leukocytes further comprises reducing platelets.

4. The method of claim 3, further comprising removing plasma and platelets from the stored OR-LRWB+PLT and adding an additive solution to the stored OR-LRWB+PLT to prepare oxygen-reduced leukopenic stored packed red blood cells (OR-LRpRBC).

5. The method of claim 1, wherein the one or more cytokines are both RANTES and eotaxin.

6. The method of claim 1 , wherein said reducing said pre-storage oxygen saturation further comprises reducing a pre-storage carbon dioxide partial pressure (pCO 2 ) to less than 60 millimeters of mercury (mmHg).

7. The method of claim 6, further comprising removing plasma from the stored OR-LRWB+PLT and adding an additive solution to the stored OR-LRWB+PLT to prepare oxygen and carbon dioxide reduced leukopenic stored packaged red blood cells (OCR-LRpRBC+PLT) containing platelets.

8. The method of claim 6, wherein said reduction of said leukocytes further comprises reducing platelets.

9. The method of claim 8, further comprising removing plasma from the stored OR-LRWB+PLT and adding an additive solution to the stored OR-LRWB+PLT to prepare oxygen and carbon dioxide reduced leukopenia packed red blood cells (OCR-LRpRBC).

10. The method of claim 1, wherein the one or more cytokines is RANTES.

11. The method of claim 1, wherein the one or more cytokines is eotaxin.

12. The method of claim 6, wherein the stored OR-LRWB+PLT has a 2,3-diphosphoglycerate (2,3-DPG) level after 15 days of storage that is higher than the initial 2,3-DPG level of the OR-LRWB+PLT at day 0.

13. The method of claim 12, wherein the 2,3-DPG level of the stored OR-LRWB+PLT after 15 days of storage is at least 80% higher than the initial 2,3-DPG level of the stored OR-LRWB+PLT at day 0.

14. The method of claim 12, wherein the 2,3-DPG level of the stored OR-LRWB+PLT after 21 days of storage is between 5 and 20 micromoles (μmol) 2,3-DPG per gram hemoglobin (gHb).

15. The method of claim 1 , wherein the stored whole blood that is oxygen-reduced and contains platelets is selected from the group consisting of: stored whole blood that is oxygen-reduced and leukocyte-reduced and contains platelets (OR-LRWB+PLT), stored packed red blood cells that are oxygen-reduced and leukocyte-reduced and contain platelets (OR-LRpRBC+PLT), stored whole blood that is oxygen- and carbon dioxide-reduced and leukocyte-reduced and contains platelets (OCR-LRWB+PLT), stored packed red blood cells that are oxygen- and carbon dioxide-reduced and leukocyte-reduced and contain platelets (OCR-LRpRBC+PLT), and any combination thereof.

16. The method of claim 1, wherein the level of the one or more cytokines in the stored OR-LRWB+PLTs after 21 days of storage is half the level of the same one or more cytokines in the conventionally stored non-OR-LRWB+PLTs.

17. The method of claim 1, wherein the levels of the one or more cytokines in the stored OR-LRWB+PLTs are relatively unchanged after 10 days of storage under anaerobic conditions.

18. The method of claim 17, wherein the levels of the one or more cytokines in the stored OR-LRWB+PLTs are relatively unchanged after storage under anaerobic conditions for 30 days.

19. The method of claim 18, wherein the levels of the one or more cytokines in the stored OR-LRWB+PLTs are relatively unchanged after 40 days of storage under anaerobic conditions.

20. The method of any one of claims 2, 4, 7, and 9, wherein the additive solution is selected from the group consisting of: Additive Solution 1 (AS-1), Additive Solution 3 (AS-3), Additive Solution 5 (AS-5), Saline-Adenine-Glucose-Mannitol (SAGM), Phosphate-Adenine-Glucose-Guanosine-Saline-Mannitol (PAGG-SM), Phosphate-Adenine-Glucose-Guanosine-Gluconate-Mannitol (PAGG-GM), Mannitol-Adenine-Phosphate (MAP), Additive Solution 7 (AS-7), Erythrosol-5 (ESOL-5), Experimental Additive Solution 61 (EAS61), Anaerobic Additive Solution 1 (OFAS1), Anaerobic Additive Solution 3 (OFAS3), and any combination thereof.

21. The method of claim 20, wherein the additive solution is AS-1.

22. The method of claim 20, wherein the additive solution is AS-3.

23. The method of claim 20, wherein the additive solution is SAGM.

24. The method of claim 10, wherein the level of RANTES in the stored OR-LRWB+PLT is half the level of RANTES in the conventionally stored non-OR-LRWB+PLT after 21 days of storage.

25. The method of claim 10, wherein the level of RANTES in the stored OR-LRWB+PLT after 40 days of storage is 25% or less of the level of RANTES in the conventionally stored non-OR-LRWB+PLT.

26. The method of claim 11, wherein the level of eotaxin in the stored OR-LRWB+PLTs after 21 days of storage is half the level of eotaxin in the conventionally stored non-OR-LRWB+PLTs.

27. The method of claim 11, wherein the level of eotaxin in the stored OR-LRWB+PLTs after 40 days of storage is 25% or less of the level of eotaxin in the conventionally stored non-OR-LRWB+PLTs.

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