Methods for preserving whole blood and composition of whole blood

Abstract

To provide methods and compositions for improved clinical outcomes for trauma patients receiving whole blood transfusion, and methods and compositions for improved clinical outcomes for blood transfusions for cancer patients.SOLUTION: A method for improving survival of a patient in need of multiple transfusions comprises providing stored red blood cells that have been reduced in oxygen (OR stored RBCs) to a patient in need thereof receiving a medical procedure.SELECTED DRAWING: Figure 1A

Description

[Technical Field]

[0001] This disclosure relates to a method for improving the quality of whole blood useful for transfusion to patients. Anaerobic storage of whole blood results in a decrease in cytokine levels and an improvement in 2,3-diphosphoglycerate (2,3-DPG) and adenosine triphosphate (ATP) levels. The improved blood composition is useful for transfusion to cancer and trauma patients. [Background technology]

[0002] When stored blood is preserved in the conventional manner, degradation associated with various preservation disorders steadily occurs, particularly hemolysis, hemoglobin degradation, and a decrease in ATP and 2,3-DPG concentrations. When transfused to a patient, the effects of this steady degradation during storage manifest, for example, as recovery within 24 hours. Due to these and other medical complications from transfusions of stored blood, various methods have been developed to minimize the impact of storage on the 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).

[0003] Numerous techniques have been developed to minimize storage damage and improve transfusion outcomes. One such technique involves the development of additives to be included during storage. Examples of this technique 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 concern additives for blood preservation and activation. For example, Rejuvesol® (available from Citra Lab LLC, Braintree, MA) is added to blood immediately before transfusion after low-temperature storage (i.e., 4°C) or before freezing for long-term storage (i.e., -80°C with glycerol). U.S. Patent No. 6,447,987 to Hess et al. concerns additives for the refrigerated storage of human red blood cells. An alternative technique is to freeze the blood to prevent storage damage. While the storage of frozen blood is known in the art, such frozen blood has limitations. U.S. Patent No. 6,413,713 to Serebrennikov et al. concerns a method for preserving blood at temperatures below 0°C. See also 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 concerns containers for blood preservation, as provided in U.S. Patent No. 4,837,047 to Sato et al.

[0004] One method proven to successfully improve blood quality and extend its usefulness is storage under oxygen-depleted and anaerobic conditions. U.S. Patents 5,624,794, 6,162,396, and 5,476,764 to Bitensky et al. concern the storage of red blood cells under oxygen-depleted conditions. U.S. Patent 5,789,151 to Bitensky et al. concerns blood preservation additives. Among the advantages of storing blood under oxygen-depleted conditions are improved ATP and 2,3-DPG levels, as well as reduced hemolysis. Furthermore, storing blood under oxygen-depleted conditions can result in lower microparticle levels, reduced loss of deformability, reduced lipid and protein oxidation, and higher post-transfusion survival rates compared to blood stored under conventional conditions.

[0005] U.S. Patent No. 6,162,396 ('396 Patent) to Bitensky et al. discloses an anaerobic storage bag for blood storage comprising an oxygen-impermeable outer layer, an oxygen-permeable erythrocyte (RBC) compatible inner layer, and an oxygen scrubber positioned between the inner and outer layers.

[0006] While the effects of oxygen depletion on packed red blood cells have been investigated, the effects of oxygen depletion on whole blood have not been reported. One reason for this lack of research on whole blood deoxygenation may be the expectation that adverse effects may occur when platelets are deprived of oxygen. More specifically, given the crucial role of platelets in the coagulation process, there is concern that impaired platelet function could lead to coagulation disorders and negatively impact clinical outcomes.

[0007] Platelet storage has been extensively studied to identify the most favorable conditions, including temperature, pH, and O2 and CO2 concentrations. The results of this research conclude that, for stored platelets to survive in recipients after transfusion, they require exposure to oxygen and storage at room temperature. Murphy and Gardner, in 1975, stated that undesirable morphological changes are associated with decreased 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 exposure to oxygen enables aerobic metabolism (oxidative phosphorylation), leading to a decrease in the rate of lactate production. At low PO2 levels, lactate production increases, consistent with the Pasteur effect. Moroff et al. stated that sustained oxygen consumption is necessary 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℃,” Vox Sanguinis 42(1):33-45 (1982). Specially adapted container systems achieve permeability to carbon dioxide and oxygen, preventing catastrophic pH drops. As shown in Kakaiya et al., “Platelet preservation in large containers,” Vox Sanguinis 46(2):111-118 (1984), maintaining platelet quality was a result of improved gas exchange conditions achieved by increasing the surface area available for gas exchange. The importance of maintaining oxygen levels during platelet storage 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 for the viability of stored platelets has been supported by the observation that lactate levels increase 5 to 8 times in oxygen-poor environments. See Kilkson et al., "Platelet metabolism during storage of platelet concentrates at 22 degrees C," Blood 64(2):406-14 (1984). Wallvik et al., "Platelet Concentrates Stored at 22℃ Need Oxygen: The Significance of Plastics in Platelet Preservation," Vox Sanguinis 45(4):303-311 (1983) reported that maintaining oxygen during the first 5 days of storage was essential for platelet preservation. Wallvik and colleagues also showed that the maximum number of platelets that can be successfully stored for 5 days is predictable based on the determination of 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 sufficient gas exchange properties, pH was maintained and ATP loss and α-granule platelet factor 4 (PF4) release were prevented. Each of the above references is incorporated herein in whole.

[0008] These findings have led to the standardization of procedures to ensure oxygenation of platelets during storage at room temperature, particularly to maximize post-transfusion viability. When platelets are stored at refrigerated temperatures, their post-transfusion viability is lost, making them unsuitable for prophylactic transfusions to cancer patients who cannot produce their own platelets. On the other hand, platelets stored at refrigerated temperatures maintain hemostatic function when transfused to the recipient. Therefore, when administering platelets to patients with traumatic bleeding, viability is less important than hemostatic activity. We demonstrated that anaerobic storage of whole blood refrigerated for up to three weeks yields hemostatic activity consistent with conventionally refrigerated whole blood, clearly showing that platelet hemostatic activity is maintained by low-temperature storage, even when they are oxygen-deficient.

[0009] While oxygen depletion in whole blood is mentioned in the literature, the effects of anaerobic storage of whole blood are not disclosed. As previously stated, it is well established that long-term survival (more than 24 hours) of platelets (PLTs) in recipients requires not only storage at room temperature but also oxygen during storage. However, in hemorrhagic trauma resuscitation, long-term survival of PLTs is not as important as their hemostatic ability. Recently, it has become clear that patients transfused with stored whole blood or fresh whole blood, and reconstituted whole blood (a mixture of plasma, red blood cells, and platelets), have significantly lower post-traumatic mortality. We have recently discovered that low-temperature storage enables anaerobic storage of PLTs and also provides the known benefits of anaerobically stored RBCs observed in packed red blood cells in whole blood. More specifically, deoxygenated whole blood provides improved 2,3-DPG levels while unexpectedly retaining coagulation ability without causing adverse effects. Over the period of storage, the deformability of RBCs is maintained under deoxygenated conditions.

[0010] As suggested by the accumulation of lipid peroxidation markers such as isoprostane, oxidative damage during storage has been considered a major cause of membrane damage in packed red blood cells (pRBCs). Increased cytokine levels during storage may also contribute to the development of storage defects that have potential clinical significance regarding the negative outcomes of transfusions.

[0011] Certain patient populations are more susceptible to the effects of storage disorders than other populations. These more susceptible populations include, non-limiting examples, trauma patients and cancer patients. Adverse clinical outcomes are associated with the accumulation of biologic response modifiers (BRMs), including cytokines, that mediate inflammation, regulate cell proliferation, regulate angiogenesis, and modulate T cell function. Examples of these BRMs include interleukin-17 (IL-17), eotaxin (CCL11), basic FGF (bFGF), macrophage inflammatory protein 1a (MIP-1a), monocyte chemotactic protein 1 (MCP-1), platelet-derived growth factor (PDGF), tumor necrosis factor α (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(Suppl3):10A (2009). Furthermore, the accumulation of cytokines during blood storage has been observed, and these accumulated cytokines may be associated with negative outcomes when administered to cancer patients perioperatively. 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). There is a need for blood storage methods that result in reduced levels of BRM and cytokines, thereby improving patient outcomes.

[0012] Traumatic injury accounts for 30% of lost life in the United States, surpassing cancer (16% of survival years) and heart disease (12%). Trauma is the leading cause of death among patients aged 1 to 46 years. Death due to bleeding often occurs within 24 hours of the traumatic injury, but premature death due to massive bleeding (within 3 to 6 hours) is avoidable with prompt and appropriate treatment.

[0013] Damage Control Resuscitation (DCR) protocols describe the concept of using balanced ratios of blood components. DCR is rapidly becoming the standard for controlling hemostasis and recovering from shock in trauma patients experiencing rapid bleeding. In civilian settings, current blood storage operations do not include whole blood stockpiles, so DCR is performed by sequential transfusions of separate components (RBCs, plasma, and platelets) so that the blood is "reconstituted" within the recipient. Earlier this year, a large randomized controlled trial (RCT), Pragmatic Randomized Optimal Platelet and Plasma Ratios (PROPPR), concluded, comparing the effectiveness of transfusing "reconstituted blood" in 1:1:1 unit ratios (plasma, platelets, and RBCs) and 1:1:2 ratios to trauma patients receiving massive transfusions. Mass transfusion kits combining packaged blood products consisting of thawed fresh frozen plasma (FFP), platelets, and RBCs in 1:1:1 ratios are now readily available in major trauma centers.

[0014] Recent studies have shown that whole blood is superior in controlling bleeding and recovering from shock in patients experiencing severe bleeding, particularly in cases of life-threatening hemorrhage. The 2015 proceedings of the NHLBI State of the Science in Transfusion Medicine Symposium prioritized research on whole blood for patients with severe bleeding. Similarly, the THOR Network, an international organization focused on damage control resuscitation, has prioritized comparative studies of the efficacy and safety of whole blood and its components for hemorrhagic shock. Because modern blood banks do not routinely supply whole blood, over 80% of the Level 1 trauma centers surveyed are attempting to mimic the hemostatic shock recovery properties required for massive transfusion protocols using plasma, platelet, and red blood cell units in 1:1:1 to 1:1:2 ratios for both traumatic and non-traumatic life-threatening bleeding cases. Providing all three blood components quickly and safely physically is challenging, especially considering the need to thaw plasma at the center when thawed plasma stock is not readily available. Recent data also shows that storing whole blood at 4°C for up to 14 days maintains better platelet function and overall hemostatic effect than storing it at 22°C.

[0015] In addition to the need for blood banks to provide whole blood for use in specific patient populations, the ability to preserve valuable blood resources is crucial. Blood banks, in particular, are failing to utilize precious and scarce resources because they typically discard whole blood stocks after two weeks (despite FDA regulations allowing for longer usable shelf lives). The ability to maximize the value of such blood resources is especially useful for smaller hospitals acting as Level III and Level IV trauma centers, where oxygen-depleted hemostatic whole blood products can be maintained under anaerobic conditions and then processed for concentrated red blood cells. This specification provides improved whole blood quality for use in trauma patients and also offers a further source of concentrated red blood cells with improved properties and less storage degradation. This specification overcomes concerns regarding the waste of valuable type O RH- RBCS (typically used in whole blood transfusions). Thus, anaerobic RBCs can be obtained from oxygen-depleted whole blood and reused in oxygen-depleted RBC units suitable for storage for up to six weeks. Where provided herein, deoxygenated packed red blood cells may be obtained from unused oxygen-depleted whole blood that has been used for transfusion or stored for later use under anaerobic conditions. [Overview of the Initiative]

[0016] This disclosure provides and includes a method for improving the survival of patients requiring frequent blood transfusions, comprising providing oxygen-reduced stored red blood cells to a patient undergoing medical treatment who requires them.

[0017] This disclosure provides and includes a method for improving the survival of cancer patients in need after perioperative transfusion, comprising providing oxygen-reduced stored red blood cells to cancer patients undergoing surgical procedures who require them.

[0018] This disclosure provides and includes a method for reducing cancer-promoting cytokines in stored blood, comprising: taking blood in an anticoagulant solution; reducing the number of white blood cells from the taken blood; reducing the oxygen saturation (SO2) to 30% or less before storage; reducing the partial pressure of carbon dioxide to 60 mmHg or less before storage; and storing the oxygen and carbon dioxide-reduced blood under anaerobic conditions, wherein the method includes depleting oxygen from the blood before storage.

[0019] This disclosure provides a blood composition for transfusion to trauma patients requiring it, comprising deoxygenated leukocyte-reduced whole blood in an anticoagulant solution, having a pre-storage oxygen saturation (SO2) of 20% or less and a pre-storage carbon dioxide partial pressure of less than 60 mmHg, wherein the deoxygenated leukocyte-reduced whole blood has a 2,3-DPG level higher than the initial 2,3-DPG level of the deoxygenated leukocyte-reduced blood on day 15.

[0020] This disclosure provides and includes a method for reducing an inflammatory response in a patient receiving a blood transfusion, comprising transfusing an oxygen-depleted blood product to a patient in need thereof, wherein the oxygen-depleted whole blood has low levels of inflammatory cytokines after storage under anaerobic conditions.

[0021] This disclosure provides and includes a method for reducing the immune response in a patient receiving a blood transfusion, comprising transfusing an oxygen-depleted blood product to a patient in need thereof, wherein the oxygen-depleted blood product has low levels of cytokines after storage under anaerobic conditions. In embodiments of this disclosure, the immune response is either immunomodulatory or immunosuppressive. In other embodiments, the immune response is, for example, activation, including inflammation.

[0022] This disclosure provides and includes a method for improving oxygen perfusion in a patient in need, comprising transfusing an oxygen-depleted blood product to the patient in need, wherein the oxygen-depleted blood product has a higher RBC deformability compared to conventionally stored blood products.

[0023] The present disclosure provides a method for managing a blood bank, including maintaining an inventory of blood units containing oxygen-reduced whole blood and an anticoagulant, or oxygen-reduced leukocyte-reduced whole blood and an anticoagulant, providing one or more of the blood units from the inventory for patient treatment, and preparing oxygen-reduced blood units with separated components by reusing the blood units from the inventory. The present disclosure further provides for the use of reused blood units for preparing reconstituted blood units for the treatment of trauma patients requiring massive transfusion.

[0024] The present disclosure provides a method for supplying blood products for transfusion medicine, including depleting oxygen or oxygen and carbon dioxide from whole blood to prepare oxygen-reduced or oxygen and carbon dioxide-reduced whole blood, storing the oxygen-reduced or oxygen and carbon dioxide-reduced whole blood for a certain period, and providing the stored blood to patients who need it, or storing the oxygen-reduced or oxygen and carbon dioxide-reduced whole blood for a period and preparing oxygen-reduced or oxygen and carbon dioxide-reduced packed red blood cells.

[0025] The present disclosure is disclosed with reference to the accompanying drawings.

Brief Description of the Drawings

[0026] [Figure 1A] Figures 1A - 1D are graphs showing the results of cytokine measurements, showing the decrease in eotaxin (1A) and RANTES (1B) levels in packed red blood cells stored anaerobically. Figure 1C shows the decrease in cell-free hemoglobin levels compared to aerobically stored packed red blood cells. Figure 1D shows the decrease in isoprostane levels in packed red blood cells stored anaerobically. Dashed line = aerobically stored blood, solid line = anaerobically stored blood. [Figure 1B]Figures 1A-1D are graphs showing the results of cytokine measurements, illustrating the decrease in eotaxin (1A) and rantes (1B) levels in anaerobically preserved packed red blood cells. Figure 1C shows the decrease in cell-free hemoglobin levels compared to aerobically preserved packed red blood cells. Figure 1D shows the decrease in isoprostane levels in anaerobically preserved packed red blood cells. Dashed lines represent aerobically preserved blood, and solid lines represent anaerobically preserved blood. [Figure 1C] Figures 1A-1D are graphs showing the results of cytokine measurements, illustrating the decrease in eotaxin (1A) and rantes (1B) levels in anaerobically preserved packed red blood cells. Figure 1C shows the decrease in cell-free hemoglobin levels compared to aerobically preserved packed red blood cells. Figure 1D shows the decrease in isoprostane levels in anaerobically preserved packed red blood cells. Dashed lines represent aerobically preserved blood, and solid lines represent anaerobically preserved blood. [Figure 1D] Figures 1A-1D are graphs showing the results of cytokine measurements, illustrating the decrease in eotaxin (1A) and rantes (1B) levels in anaerobically preserved packed red blood cells. Figure 1C shows the decrease in cell-free hemoglobin levels compared to aerobically preserved packed red blood cells. Figure 1D shows the decrease in isoprostane levels in anaerobically preserved packed red blood cells. Dashed lines represent aerobically preserved blood, and solid lines represent anaerobically preserved blood.

[0027] [Figure 2A]Figures 2A-2G are graphs showing the results of two experiments according to this disclosure, comparing the storage of leukocyte-reduced whole blood (LRWB / CPD) collected in anticoagulant solution CPD under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPD over a period of 21 days. Figure 2A shows the level of 2,3-DPG. Figure 2B shows the level of ATP. Figure 2C shows the pH. Figure 2D shows the platelet count. Figure 2E shows the potassium level. Figure 2F shows the data from Figure 2A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 2G shows the data from Figure 2B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c68 / 80 is conventionally stored blood with an initial oxygen saturation of 68% and a CO2 partial pressure of 80 mmHg; Sample c50 / 94 is conventionally stored blood with an initial oxygen saturation of 50% and a CO2 partial pressure of 94 mmHg; Sample sc91 / 75 is conventionally stored blood with an initial oxygen saturation of 91% and a CO2 partial pressure of 75 mmHg; Sample sc69 / 87 is conventionally stored blood with an initial oxygen saturation of 69% and a CO2 partial pressure of 87 mmHg; Sample tc5 / 78 is conventionally stored blood with an initial oxygen saturation of 5% Sample tc7 / 64 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 78 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T4 / 26 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 4% and a CO2 partial pressure of 26 mmHg. [Figure 2B]Figures 2A-2G are graphs showing the results of two experiments according to this disclosure, comparing the storage of leukocyte-reduced whole blood (LRWB / CPD) collected in anticoagulant solution CPD under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPD over a period of 21 days. Figure 2A shows the level of 2,3-DPG. Figure 2B shows the level of ATP. Figure 2C shows the pH. Figure 2D shows the platelet count. Figure 2E shows the potassium level. Figure 2F shows the data from Figure 2A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 2G shows the data from Figure 2B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c68 / 80 is conventionally stored blood with an initial oxygen saturation of 68% and a CO2 partial pressure of 80 mmHg; Sample c50 / 94 is conventionally stored blood with an initial oxygen saturation of 50% and a CO2 partial pressure of 94 mmHg; Sample sc91 / 75 is conventionally stored blood with an initial oxygen saturation of 91% and a CO2 partial pressure of 75 mmHg; Sample sc69 / 87 is conventionally stored blood with an initial oxygen saturation of 69% and a CO2 partial pressure of 87 mmHg; Sample tc5 / 78 is conventionally stored blood with an initial oxygen saturation of 5% Sample tc7 / 64 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 78 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T4 / 26 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 4% and a CO2 partial pressure of 26 mmHg. [Figure 2C]Figures 2A-2G are graphs showing the results of two experiments according to this disclosure, comparing the storage of leukocyte-reduced whole blood (LRWB / CPD) collected in anticoagulant solution CPD under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPD over a period of 21 days. Figure 2A shows the level of 2,3-DPG. Figure 2B shows the level of ATP. Figure 2C shows the pH. Figure 2D shows the platelet count. Figure 2E shows the potassium level. Figure 2F shows the data from Figure 2A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 2G shows the data from Figure 2B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c68 / 80 is conventionally stored blood with an initial oxygen saturation of 68% and a CO2 partial pressure of 80 mmHg; Sample c50 / 94 is conventionally stored blood with an initial oxygen saturation of 50% and a CO2 partial pressure of 94 mmHg; Sample sc91 / 75 is conventionally stored blood with an initial oxygen saturation of 91% and a CO2 partial pressure of 75 mmHg; Sample sc69 / 87 is conventionally stored blood with an initial oxygen saturation of 69% and a CO2 partial pressure of 87 mmHg; Sample tc5 / 78 is conventionally stored blood with an initial oxygen saturation of 5% Sample tc7 / 64 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 78 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T4 / 26 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 4% and a CO2 partial pressure of 26 mmHg. [Figure 2D]Figures 2A-2G are graphs showing the results of two experiments according to this disclosure, comparing the storage of leukocyte-reduced whole blood (LRWB / CPD) collected in anticoagulant solution CPD under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPD over a period of 21 days. Figure 2A shows the level of 2,3-DPG. Figure 2B shows the level of ATP. Figure 2C shows the pH. Figure 2D shows the platelet count. Figure 2E shows the potassium level. Figure 2F shows the data from Figure 2A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 2G shows the data from Figure 2B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c68 / 80 is conventionally stored blood with an initial oxygen saturation of 68% and a CO2 partial pressure of 80 mmHg; Sample c50 / 94 is conventionally stored blood with an initial oxygen saturation of 50% and a CO2 partial pressure of 94 mmHg; Sample sc91 / 75 is conventionally stored blood with an initial oxygen saturation of 91% and a CO2 partial pressure of 75 mmHg; Sample sc69 / 87 is conventionally stored blood with an initial oxygen saturation of 69% and a CO2 partial pressure of 87 mmHg; Sample tc5 / 78 is conventionally stored blood with an initial oxygen saturation of 5% Sample tc7 / 64 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 78 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T4 / 26 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 4% and a CO2 partial pressure of 26 mmHg. [Figure 2E]Figures 2A-2G are graphs showing the results of two experiments according to this disclosure, comparing the storage of leukocyte-reduced whole blood (LRWB / CPD) collected in anticoagulant solution CPD under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPD over a period of 21 days. Figure 2A shows the level of 2,3-DPG. Figure 2B shows the level of ATP. Figure 2C shows the pH. Figure 2D shows the platelet count. Figure 2E shows the potassium level. Figure 2F shows the data from Figure 2A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 2G shows the data from Figure 2B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c68 / 80 is conventionally stored blood with an initial oxygen saturation of 68% and a CO2 partial pressure of 80 mmHg; Sample c50 / 94 is conventionally stored blood with an initial oxygen saturation of 50% and a CO2 partial pressure of 94 mmHg; Sample sc91 / 75 is conventionally stored blood with an initial oxygen saturation of 91% and a CO2 partial pressure of 75 mmHg; Sample sc69 / 87 is conventionally stored blood with an initial oxygen saturation of 69% and a CO2 partial pressure of 87 mmHg; Sample tc5 / 78 is conventionally stored blood with an initial oxygen saturation of 5% Sample tc7 / 64 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 78 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T4 / 26 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 4% and a CO2 partial pressure of 26 mmHg. [Figure 2F]Figures 2A-2G are graphs showing the results of two experiments according to this disclosure, comparing the storage of leukocyte-reduced whole blood (LRWB / CPD) collected in anticoagulant solution CPD under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPD over a period of 21 days. Figure 2A shows the level of 2,3-DPG. Figure 2B shows the level of ATP. Figure 2C shows the pH. Figure 2D shows the platelet count. Figure 2E shows the potassium level. Figure 2F shows the data from Figure 2A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 2G shows the data from Figure 2B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c68 / 80 is conventionally stored blood with an initial oxygen saturation of 68% and a CO2 partial pressure of 80 mmHg; Sample c50 / 94 is conventionally stored blood with an initial oxygen saturation of 50% and a CO2 partial pressure of 94 mmHg; Sample sc91 / 75 is conventionally stored blood with an initial oxygen saturation of 91% and a CO2 partial pressure of 75 mmHg; Sample sc69 / 87 is conventionally stored blood with an initial oxygen saturation of 69% and a CO2 partial pressure of 87 mmHg; Sample tc5 / 78 is conventionally stored blood with an initial oxygen saturation of 5% Sample tc7 / 64 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 78 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T4 / 26 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 4% and a CO2 partial pressure of 26 mmHg. [Figure 2G]Figures 2A-2G are graphs showing the results of two experiments according to this disclosure, comparing the storage of leukocyte-reduced whole blood (LRWB / CPD) collected in anticoagulant solution CPD under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPD over a period of 21 days. Figure 2A shows the level of 2,3-DPG. Figure 2B shows the level of ATP. Figure 2C shows the pH. Figure 2D shows the platelet count. Figure 2E shows the potassium level. Figure 2F shows the data from Figure 2A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 2G shows the data from Figure 2B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c68 / 80 is conventionally stored blood with an initial oxygen saturation of 68% and a CO2 partial pressure of 80 mmHg; Sample c50 / 94 is conventionally stored blood with an initial oxygen saturation of 50% and a CO2 partial pressure of 94 mmHg; Sample sc91 / 75 is conventionally stored blood with an initial oxygen saturation of 91% and a CO2 partial pressure of 75 mmHg; Sample sc69 / 87 is conventionally stored blood with an initial oxygen saturation of 69% and a CO2 partial pressure of 87 mmHg; Sample tc5 / 78 is conventionally stored blood with an initial oxygen saturation of 5% Sample tc7 / 64 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 78 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T4 / 26 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 4% and a CO2 partial pressure of 26 mmHg.

[0028] [Figure 3A]Figures 3A-3D are graphs showing the results of two experiments according to this disclosure, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in anticoagulant solution CPDA1 under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 3A shows the level of 2,3-DPG. Figure 3B shows the level of ATP. Figure 3C shows the data from Figure 3A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 3D shows the data from Figure 3B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c32 / 98 is conventionally stored blood with an initial oxygen saturation of 32% and a CO2 partial pressure of 98 mmHg; Sample c56 / 86 is conventionally stored blood with an initial oxygen saturation of 56% and a CO2 partial pressure of 86 mmHg; Sample sc59 / 95 is conventionally stored blood with an initial oxygen saturation of 59% and a CO2 partial pressure of 95 mmHg; Sample sc82 / 84 is conventionally stored blood with an initial oxygen saturation of 82% and a CO2 partial pressure of 84 mmHg; Sample tc7 / 80 is conventionally stored blood with an initial oxygen saturation of 7% Sample tc6 / 77 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 6% and a CO2 partial pressure of 80 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T7 / 23 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 23 mmHg. [Figure 3B]Figures 3A-3D are graphs showing the results of two experiments according to this disclosure, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in anticoagulant solution CPDA1 under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 3A shows the level of 2,3-DPG. Figure 3B shows the level of ATP. Figure 3C shows the data from Figure 3A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 3D shows the data from Figure 3B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c32 / 98 is conventionally stored blood with an initial oxygen saturation of 32% and a CO2 partial pressure of 98 mmHg; Sample c56 / 86 is conventionally stored blood with an initial oxygen saturation of 56% and a CO2 partial pressure of 86 mmHg; Sample sc59 / 95 is conventionally stored blood with an initial oxygen saturation of 59% and a CO2 partial pressure of 95 mmHg; Sample sc82 / 84 is conventionally stored blood with an initial oxygen saturation of 82% and a CO2 partial pressure of 84 mmHg; Sample tc7 / 80 is conventionally stored blood with an initial oxygen saturation of 7% Sample tc6 / 77 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 6% and a CO2 partial pressure of 80 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T7 / 23 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 23 mmHg. [Figure 3C]Figures 3A-3D are graphs showing the results of two experiments according to this disclosure, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in anticoagulant solution CPDA1 under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 3A shows the level of 2,3-DPG. Figure 3B shows the level of ATP. Figure 3C shows the data from Figure 3A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 3D shows the data from Figure 3B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c32 / 98 is conventionally stored blood with an initial oxygen saturation of 32% and a CO2 partial pressure of 98 mmHg; Sample c56 / 86 is conventionally stored blood with an initial oxygen saturation of 56% and a CO2 partial pressure of 86 mmHg; Sample sc59 / 95 is conventionally stored blood with an initial oxygen saturation of 59% and a CO2 partial pressure of 95 mmHg; Sample sc82 / 84 is conventionally stored blood with an initial oxygen saturation of 82% and a CO2 partial pressure of 84 mmHg; Sample tc7 / 80 is conventionally stored blood with an initial oxygen saturation of 7% Sample tc6 / 77 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 6% and a CO2 partial pressure of 80 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T7 / 23 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 23 mmHg. [Figure 3D]Figures 3A-3D are graphs showing the results of two experiments according to this disclosure, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in anticoagulant solution CPDA1 under oxygen reduction, oxygen and carbon dioxide reduction, and conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 3A shows the level of 2,3-DPG. Figure 3B shows the level of ATP. Figure 3C shows the data from Figure 3A plotted again in comparison to the level of 2,3-DPG on day 0 (T0). Figure 3D shows the data from Figure 3B plotted again in comparison to the level of ATP on day 0 (T0). Legend: Sample c32 / 98 is conventionally stored blood with an initial oxygen saturation of 32% and a CO2 partial pressure of 98 mmHg; Sample c56 / 86 is conventionally stored blood with an initial oxygen saturation of 56% and a CO2 partial pressure of 86 mmHg; Sample sc59 / 95 is conventionally stored blood with an initial oxygen saturation of 59% and a CO2 partial pressure of 95 mmHg; Sample sc82 / 84 is conventionally stored blood with an initial oxygen saturation of 82% and a CO2 partial pressure of 84 mmHg; Sample tc7 / 80 is conventionally stored blood with an initial oxygen saturation of 7% Sample tc6 / 77 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 6% and a CO2 partial pressure of 80 mmHg; Sample T5 / 28 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 5% and a CO2 partial pressure of 28 mmHg; Sample T7 / 23 is oxygen-depleted, anaerobically stored blood with an initial oxygen saturation of 7% and a CO2 partial pressure of 23 mmHg.

[0029] [Figure 4A]Figures 4A-4C are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in anticoagulant solution CPDA1 under oxygen reduction (OR), oxygen and carbon dioxide reduction (OCR), and conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 4A shows the levels of ATP in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1. Figure 4B shows the levels of 2,3-DPG in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1. Figure 4C shows the percentage of hemolysis in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1. In the graphs shown in Figures 4A to 4C, the dotted line represents OR-LRWB / CPDA1 stored blood, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents blood stored in the conventional manner. [Figure 4B] Figures 4A-4C are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in anticoagulant solution CPDA1 under oxygen reduction (OR), oxygen and carbon dioxide reduction (OCR), and conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 4A shows the levels of ATP in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1. Figure 4B shows the levels of 2,3-DPG in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1. Figure 4C shows the percentage of hemolysis in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1. In the graphs shown in Figures 4A to 4C, the dotted line represents OR-LRWB / CPDA1 stored blood, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents blood stored in the conventional manner. [Figure 4C]Figures 4A-4C are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in anticoagulant solution CPDA1 under oxygen reduction (OR), oxygen and carbon dioxide reduction (OCR), and conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 4A shows the levels of ATP in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1. Figure 4B shows the levels of 2,3-DPG in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1. Figure 4C shows the percentage of hemolysis in OR-LRWB / CPDA1, OCR-LRWB / CPDA1, and conventionally stored LRWB / CPDA1. In the graphs shown in Figures 4A to 4C, the dotted line represents OR-LRWB / CPDA1 stored blood, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents blood stored in the conventional manner.

[0030] [Figure 5A] Figures 5A-5D are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under reduced oxygen and carbon dioxide conditions with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 5A shows the activated partial thrombin time (aPTT) in seconds for OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5B shows the prothrombin time (PT) in seconds for OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5C shows the fibrinogen levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5D shows the D-dimer levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph representing plasma coagulation parameters, the dashed line represents ORC-LRWB / CPDA1 stored blood, while the solid line represents blood stored in the conventional manner. [Figure 5B]Figures 5A-5D are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under reduced oxygen and carbon dioxide conditions with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 5A shows the activated partial thrombin time (aPTT) in seconds for OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5B shows the prothrombin time (PT) in seconds for OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5C shows the fibrinogen levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5D shows the D-dimer levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph representing plasma coagulation parameters, the dashed line represents ORC-LRWB / CPDA1 stored blood, while the solid line represents blood stored in the conventional manner. [Figure 5C] Figures 5A-5D are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under reduced oxygen and carbon dioxide conditions with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 5A shows the activated partial thrombin time (aPTT) in seconds for OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5B shows the prothrombin time (PT) in seconds for OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5C shows the fibrinogen levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5D shows the D-dimer levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph representing plasma coagulation parameters, the dashed line represents ORC-LRWB / CPDA1 stored blood, while the solid line represents blood stored in the conventional manner. [Figure 5D]Figures 5A-5D are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under reduced oxygen and carbon dioxide conditions with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 5A shows the activated partial thrombin time (aPTT) in seconds for OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5B shows the prothrombin time (PT) in seconds for OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5C shows the fibrinogen levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 5D shows the D-dimer levels in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph representing plasma coagulation parameters, the dashed line represents ORC-LRWB / CPDA1 stored blood, while the solid line represents blood stored in the conventional manner.

[0031] [Figure 6A] Figures 6A-6E are graphs showing the results of the experiment described herein, comparing leukopenic whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under reduced oxygen and carbon dioxide conditions with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 6A shows the levels of factor V in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6B shows the levels of factor VIII in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6C shows the protein C activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6D shows the protein S activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6E shows the levels of von Willebrand factor (vWF) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph representing plasma coagulation factors, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents conventionally stored blood. [Figure 6B] Figures 6A-6E are graphs showing the results of the experiment described herein, comparing leukopenic whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under reduced oxygen and carbon dioxide conditions with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 6A shows the levels of factor V in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6B shows the levels of factor VIII in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6C shows the protein C activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6D shows the protein S activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6E shows the levels of von Willebrand factor (vWF) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph representing plasma coagulation factors, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents conventionally stored blood. [Figure 6C]Figures 6A-6E are graphs showing the results of the experiment described herein, comparing leukopenic whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under reduced oxygen and carbon dioxide conditions with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 6A shows the levels of factor V in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6B shows the levels of factor VIII in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6C shows the protein C activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6D shows the protein S activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6E shows the levels of von Willebrand factor (vWF) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph representing plasma coagulation factors, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents conventionally stored blood. [Figure 6D] Figures 6A-6E are graphs showing the results of the experiment described herein, comparing leukopenic whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under reduced oxygen and carbon dioxide conditions with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 6A shows the levels of factor V in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6B shows the levels of factor VIII in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6C shows the protein C activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6D shows the protein S activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6E shows the levels of von Willebrand factor (vWF) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph representing plasma coagulation factors, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents conventionally stored blood. [Figure 6E] Figures 6A-6E are graphs showing the results of the experiment described herein, comparing leukopenic whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under reduced oxygen and carbon dioxide conditions with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 6A shows the levels of factor V in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6B shows the levels of factor VIII in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6C shows the protein C activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6D shows the protein S activity in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 6E shows the levels of von Willebrand factor (vWF) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph representing plasma coagulation factors, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents conventionally stored blood.

[0032] [Figure 7A]Figures 7A-7D are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under oxygen and carbon dioxide reduction (OCR) with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 7A shows the rate of fibrin deposition and crosslinking (TEG angle) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7B shows a comparison of blood pharmacokinetics (TEG K) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7C shows the maximum amplitude in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7D shows the reaction time in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph showing thromboelastography (TEG) parameters, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents blood stored in the conventional manner. [Figure 7B] Figures 7A-7D are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under oxygen and carbon dioxide reduction (OCR) with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 7A shows the rate of fibrin deposition and crosslinking (TEG angle) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7B shows a comparison of blood pharmacokinetics (TEG K) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7C shows the maximum amplitude in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7D shows the reaction time in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph showing thromboelastography (TEG) parameters, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents blood stored in the conventional manner. [Figure 7C]Figures 7A-7D are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under oxygen and carbon dioxide reduction (OCR) with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 7A shows the rate of fibrin deposition and crosslinking (TEG angle) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7B shows a comparison of blood pharmacokinetics (TEG K) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7C shows the maximum amplitude in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7D shows the reaction time in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph showing thromboelastography (TEG) parameters, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents blood stored in the conventional manner. [Figure 7D] Figures 7A-7D are graphs showing the results of the experiment described herein, comparing leukocyte-reduced whole blood (LRWB / CPDA1) collected in an anticoagulant solution CPDA1 under oxygen and carbon dioxide reduction (OCR) with conventionally stored LRWB / CPDA1 over a period of 21 days. Figure 7A shows the rate of fibrin deposition and crosslinking (TEG angle) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7B shows a comparison of blood pharmacokinetics (TEG K) in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7C shows the maximum amplitude in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. Figure 7D shows the reaction time in OCR-LRWB / CPDA1 and conventionally stored LRWB / CPDA1. In the graph showing thromboelastography (TEG) parameters, the dashed line represents ORC-LRWB / CPDA1 stored blood, and the solid line represents blood stored in the conventional manner. [Modes for carrying out the invention]

[0033] Unless otherwise defined, scientific and technical terms used herein have the same meanings as those generally understood by those skilled in the art. Those skilled in the art will recognize that many methods may be used in the practice of this disclosure. Indeed, this disclosure does not limit the methods and materials described herein. Any references cited herein are incorporated herein by reference in their entirety. For the purposes of this disclosure, the following terms are defined below:

[0034] As used herein, the term “patient” includes any person who requires medical treatment to receive blood products.

[0035] As used herein, the term “frequent transfusion” refers to a patient receiving more than 195 units of blood. In another embodiment, frequent transfusion is when a patient receives at least 1 × 10⁶ units of blood. 5 This may include receiving mL of blood. In another embodiment, frequent transfusions may involve the patient receiving 1 to 1 × 10⁶ mL of blood. 5 This involves receiving mL of blood. In another embodiment, frequent transfusions involve the patient receiving 1 × 10 mL of blood. 4 ~1 × 10 5 This includes receiving mL of blood.

[0036] As used herein, the term “blood” refers to whole blood, leukocyte-reduced RBCs, thrombocytopenic RBCs, and leukocyte and thrombocytopenic RBCs. The term “blood” further 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 blood may vary depending on the stage of the collection process, starting at 37°C, which is normal body temperature at the time of collection, rapidly decreasing to about 30°C as soon as the blood leaves the patient’s body, and if left unprocessed, further decreasing to room temperature over about 6 hours, and finally being refrigerated to about 4°C–6°C.

[0037] As used herein, “blood product” includes isolated platelets, plasma, or leukocytes.

[0038] As used herein, “recovered blood products” includes isolated platelets, plasma, or leukocytes collected from a donor.

[0039] As used herein, “recovered blood” includes whole blood and red blood cells collected from a donor and previously stored under oxygen-reduced conditions. In some embodiments of this disclosure, blood suitable for use in this method 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), oxygen-reduced-leukocyte-reduced packed red blood cells (OR-LRpRBC+PLT), oxygen and carbon dioxide-reduced-leukocyte-reduced packed red blood cells (OCR-LRpRBC), or oxygen and carbon dioxide-reduced-leukocyte-reduced packed red blood cells (OCR-LRpRBC+PLT) obtained from oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood (OCR-LRpRBC+PLT) after being stored for at least one week. In another embodiment, blood suitable for use in this method is stored for up to 42 days. In another embodiment, blood suitable for use in this method is stored for up to 56 days. In another embodiment, blood suitable for use in this method is stored for up to 64 days.

[0040] As used herein, a method for obtaining a “component-separated blood product” includes obtaining reusable blood from a blood bank's inventory and separating it into platelets, plasma, and leukocytes. Blood suitable for use in this method includes oxygen-depleted whole blood with an anticoagulant and oxygen-depleted-leukocyte-depleted whole blood with an anticoagulant. In one aspect of this disclosure, the component-separated oxygen-depleted blood is stored for up to six weeks. In another aspect, the component-separated oxygen-depleted blood includes an additive. In a particular aspect, the additive may be AS-1. In a particular aspect, the additive is AS-3 (Nutricel®). In a particular aspect, the additive is AS-5. In a particular aspect, the additive is SAGM. In a particular aspect, the additive is PAGG-SM. In a particular aspect, the additive is PAGG-GM. In a particular aspect, the additive is MAP. In a particular aspect, the additive is SOLX. In a particular aspect, the additive is ESOL. In a particular aspect, the additive is EAS61. In a particular embodiment, the additive is OFAS1. In a particular embodiment, the additive is OFAS3. In a particular embodiment, the additive is a combination of AS-1, AS-3 (Nutricel®), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, SOLX, ESOL, EAS61, OFAS1, and OFAS3, either alone or in combination.

[0041] As used herein, “reconstituted WB” includes providing platelets, RBCs, and plasma to a patient in parallel during a transfusion.

[0042] As used herein, “induced WB” includes oxygen-reduced whole blood and oxygen and carbon dioxide-reduced whole blood.

[0043] As used herein, “stored red blood cells” includes oxygen-depleted red blood cells or oxygen and carbon dioxide-depleted red blood cells stored at 1-6°C. In one embodiment, stored red blood cells include red blood cells (RBCs) present in whole blood. In another embodiment, stored red blood cells include RBCs present in leukopenic whole blood. In yet another embodiment, stored red blood cells include red blood cells (RBCs) present in leukopenic RBCs. In a further embodiment, stored red blood cells include red blood cells (RBCs) present in platelet-depleted RBCs. In yet another embodiment, stored red blood cells include red blood cells (RBCs) present in both leukopenic and platelet-depleted RBCs.

[0044] As used herein, "whole blood" includes white blood cells (WBCs), platelets suspended in plasma, electrolytes, hormones, vitamins, antibodies, etc. Whole blood typically contains 4.5 to 11.0 to 10 white blood cells. 9 A normal RBC at sea level is present in the range of cells / L, with a density of 4.6–6.2 × 10⁴ in males. 12 / L, for women: 4.2~5.4 x 10 12 The value is / L. Normal hematocrit, or blood cell volume, is approximately 40-54% in men and 38-47% in women. Platelet count is usually 150-450 × 10⁶ in both men and women. 9 The volume is / L. Whole blood is collected from a donor and is usually combined with an anticoagulant. The collected whole blood is initially at approximately 37°C and is rapidly cooled to approximately 30°C during and immediately after collection, but is gradually cooled to ambient temperature over approximately 6 hours. The whole blood may be processed at the time of collection according to the method of this disclosure, starting at 30–37°C or at room temperature (typically about 25°C). As used herein, a “unit” of blood, including the anticoagulant, is approximately 450–500 ml. Suitable anticoagulants include CPD, CPDA1, ACD, and ACD-A. As used herein, “collection time” (Tc) is the time at which blood is collected from the patient.

[0045] As used herein, “red blood cells” (RBCs), stored red blood cells, oxygen-reduced red blood cells, and oxygen and carbon dioxide-reduced red blood cells include RBCs present in whole blood, leukocyte-reduced RBCs, platelet-reduced RBCs, leukocyte and platelet-reduced RBCs, and packed red blood cells (pRBCs). Human red blood cells in the body are in a dynamic state. Red blood cells contain hemoglobin, an iron-containing protein that carries oxygen throughout the body and gives blood its red color. The percentage of blood volume composed of red blood cells is called the hematocrit value. As used herein, unless otherwise limited, RBCs also include packed red blood cells (pRBCs). Packed red blood cells are prepared from whole blood using centrifugation techniques commonly known in the art. As used herein, unless otherwise indicated, the hematocrit value of pRBCs is approximately 70%. As used herein, oxygen-reduced RBCs (OR-RBCs) may include RBCs with reduced oxygen and carbon dioxide (OCR-) (OCR-RBCs).

[0046] As used herein, “leukopenic whole blood” (LRWB) typically comprises whole blood treated with an anticoagulant to reduce leukocytes and platelets, usually by filtration or centrifugation. Leukopenic whole blood has a leukocyte level reduced by at least 5 logarithmically.

[0047] As used herein, "oxygen-reduced-leukocyte-reduced whole blood" (OR-LRWB) may include oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood (OCR-LRWB).

[0048] As used herein, “platelet-containing leukocyte-reduced whole blood” (LRWB+PLT) includes oxygen-reduced (OR-) whole blood having an anticoagulant and reduced leukocytes using a platelet preservation filter. As used herein, platelet-containing oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB+PLT) may include platelet-containing oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood (OCR-LRWB+PLT).

[0049] As used herein, “leukocyte-reduced packed red blood cells” (LRpRBCs) typically include packed red blood cells having oxygen-reduced (OR-) whole blood with an anticoagulant, which has been treated to reduce leukocytes by filtration or centrifugation. As used herein, oxygen-reduced-leukocyte-reduced packed red blood cells (OR-LRpRBCs) may include oxygen and carbon dioxide-reduced-leukocyte-reduced packed red blood cells (OCR-LRpRBCs).

[0050] As used herein, “platelet-containing leukocyte-reduced packed red blood cells” (LRpRBC+PLT) includes platelet-containing packed red blood cells obtained from oxygen-reduced whole blood with an anticoagulant, which has been treated to reduce leukocytes using a platelet-preserving filter. As used herein, oxygen-reduced-leukocyte-reduced packed red blood cells containing platelets (OR-LRpRBC+PLT) may include oxygen- and carbon dioxide-reduced-leukocyte-reduced packed red blood cells containing platelets (OCR-LRpRBC+PLT).

[0051] In aspects of this disclosure, the method and composition may include adding an additive to concentrated RBCs to form a suspension. Numerous additives are known in the art. In certain aspects, the additive may be selected individually or in combination from the group consisting of AS-1, AS-3 (Nutricel®), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5, EAS61, OFAS1, and OFAS3. Additive AS-1 is disclosed in Heaton et al., "Use of Adsol preservation solution for prolonged storage of low viscosity AS-1 red blood cells," Br J Haematol., 57(3):467-78 (1984). In further aspects, the additive may have a pH of 5.0 to 9.0. In another aspect, the additive may include an antioxidant. In some aspects of this disclosure, the antioxidant may be quercetin, α-tocopherol, ascorbic acid, or an oxidase enzyme inhibitor.

[0052] As used herein, the term “approximately” refers to ±10%.

[0053] The terms "comprises," "comprising," "includes," "including," and "having," as well as their conjugations, all mean "to include, but not necessarily, ~."

[0054] The term "~naru" means "to include and be limited to ~".

[0055] The term "essentially consisting of" means that the composition, method, or structure may include those additional components, steps, and / or parts, provided that those additional components, steps, and / or parts do not substantially affect the basic and novel features of the claimed composition, method, or structure.

[0056] As used herein, the singular forms "a," "an," and "the" refer to multiple subjects unless otherwise clearly indicated by the context. For example, "a compound" or "at least one compound" may refer to multiple compounds, including mixtures thereof.

[0057] Throughout this application, various aspects of the disclosure may be presented in scope form. It should be understood that the scope form is for convenience and conciseness only and should not be interpreted as a firm limitation on the scope of the disclosure. Therefore, the scope description should be considered to specifically disclose all conceivable partial ranges and the individual numbers within those ranges. For example, a range description such as "1-6" should be considered to specifically disclose not only partial ranges such as "1-3," "1-4," "1-5," "2-4," "2-6," and "3-6," but also the individual numbers within those ranges, such as 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0058] Whenever a numerical range is indicated in this specification, it means that any number (fraction or integer) listed within that range is included. The phrases "range extending between / between" the first and second indicators, and "range from" the first indicator to / up to the second indicator, are used synonymously in this specification and mean that the first and second indicators and all decimals and integers between them are included.

[0059] As used herein, the term “method” means a method, means, techniques and procedures for accomplishing a given task, including but not limited to methods, means, techniques and procedures that are known or readily developed from known methods, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical fields.

[0060] As used herein, the term “equal” means that, when comparing measurements of oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) with measurements of conventionally stored blood processed in an equivalent manner in another way, the values ​​are within one standard deviation of each other for at least five sample sizes for each comparative measurement condition.

[0061] As used herein, the terms “higher” or “increased” mean that, when comparing measurements of oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) to OR-WB, the measurements of conventionally stored blood processed in an equivalent manner by another method are at least one standard deviation higher for at least five sample sizes for each comparative measurement condition.

[0062] As used herein, the terms “reduced” or “less” mean that, when comparing measurements of oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) to OR-WB, the measurements of conventionally stored blood processed in an equivalent manner by another method are at least one standard deviation lower for at least five sample sizes for each comparative measurement condition.

[0063] As used herein, the terms "preserved as conventional", "conventional preservation", and "conventional conditions" refer to whole blood, leukoreduced RBC, platelet-reduced RBC, leukocyte- and platelet-reduced RBC, packed red blood cells, platelet-reduced packed red blood cells, leukoreduced packed red blood cells (LRpRBC), and leukocyte- and platelet-reduced packed red blood cells preserved in an oxygen and carbon dioxide permeable container at 1-6 °C without a gas reduction step prior to preservation. In one aspect of the present disclosure, both oxygen and carbon dioxide increase over time to ambient levels in whole blood, leukoreduced RBC, platelet-reduced RBC, leukocyte- and platelet-reduced RBC, packed red blood cells, platelet-reduced packed red blood cells, leukoreduced packed red blood cells (LRpRBC), and leukocyte- and platelet-reduced packed red blood cells preserved as conventional due to the container permeability of oxygen and carbon dioxide. Although not considered conventional per se, for the purposes of the present disclosure, conventional preservation can include preservation at temperatures above 6 °C. Also, although not considered conventional per se, for the purposes of the present disclosure, conventional preservation can include preservation at freezing temperatures.

[0064] The present disclosure provides and includes methods for providing desirable characteristics to blood products for transfusion. Depletion of oxygen from packed red blood cells results in unbound cytokines, particularly RANTES (C-C motif chemokine ligand 5, CCL5) and eotaxin (C-C motif chemokine ligand 11, CCL11), as well as cell-free hemoglobin and 8-isoprostane F 2αIt was discovered that this leads to a reduction in the accumulation of 2,3-DPG. Although not constrained by theory, it is thought that RANTES and eotaxin are normally sequestered by binding to DARC (atypical chemokine receptor 1, ACKR1) and oxidative stress damage to DARC, releasing bound chemokines. Thus, the total chemokine content does not change, but the effective concentrations (e.g., free diffusion and unbound) increase, which can then affect transfused patients. As understood, the presence of these active chemokines (acting in a dose-dependent manner) can be detrimental to trauma patients and other patients receiving two or more transfusions. These findings demonstrate an unexpected benefit of anaerobic storage of blood, in addition to the desirable efficient oxygen delivery associated with elevated 2,3-DPG levels, and offer a potential reduction in some of the components of storage impairment resulting from oxidative damage to pRBCs during storage. These cytokines are known to be negatively associated with patient outcomes in several patient populations. Therefore, the finding that the accumulation of unbound cytokines can be reduced provides an improved way to treat patients susceptible to cytokine effects.

[0065] This disclosure provides and includes improving the survival of patients requiring frequent transfusions by providing oxygen-reduced stored red blood cells (OR stored RBCs) to patients undergoing medical procedures or in need thereof. While not constrained by theory, elevated cytokine levels are thought to have adverse effects on recipient patients and increase mortality. In one embodiment, stored red blood cells are oxygen-reduced (OR). In a further embodiment, stored red blood cells are oxygen-reduced and carbon dioxide-reduced (OCR). As shown in the examples, in OCR samples, ATP levels were reduced and maintained at lower levels for at least 15 days, whereas in OR samples, ATP levels were elevated compared to conventionally stored samples (see Figure 4A). As shown in the examples, in OCR samples, 2,3-DPG levels were elevated and maintained at higher levels for at least 15 days, whereas in OR samples, 2,3-DPG levels increased over conventional storage but not as high as the 2,3-DPG levels in OCR samples (see Figure 4B). Furthermore, as shown in the examples, hemolysis was comparable in OR samples, OCR samples, and conventionally stored samples.

[0066] In one aspect of this disclosure, the cytokine comprises monocyte chemotactic protein-1 (MCP-1). In another aspect, the cytokine comprises activated regulated normal T cell expression and secretion (RANTES). In yet another aspect, the cytokine comprises angiogenin. In yet another aspect of this disclosure, the cytokine comprises tumor necrosis factor-α (TNF-α). In yet another aspect, the cytokine comprises epidermal growth factor (EGF). In a further aspect, the cytokine comprises platelet-derived growth factor (PDGF).

[0067] In one aspect of this disclosure, the level of RANTES factor is less than 500 pg / ml after 21 days under OR conditions. In another aspect, the level of RANTES factor is less than 400 pg / ml after 21 days under OR conditions. In yet another aspect, the level of RANTES factor is less than 300 pg / ml after 21 days under OR conditions. In yet another aspect, the level of RANTES factor is greater than 100 pg / ml after 21 days under OR conditions. In a further aspect, the level of RANTES factor is between 0 and 300 pg / ml after 21 days under OR conditions.

[0068] In one aspect of this disclosure, the level of eotaxin factor is less than 150 pg / ml after 21 days under OR conditions. In another aspect, the level of eotaxin factor is less than 100 pg / ml after 21 days under OR conditions. In yet another aspect, the level of eotaxin factor is between 0 and 100 pg / ml after 21 days under OR conditions. In yet another aspect, the level of eotaxin factor is preferably 100 pg / ml after 21 days under OR conditions. In yet another aspect, the level of eotaxin factor is greater than 100 pg / ml after 21 days under OR conditions. In a further aspect, the level of eotaxin factor is between 0 and 300 pg / ml after 21 days under OR conditions.

[0069] In an embodiment of this disclosure, OR-preserved RBCs are selected from the group consisting of oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen-depleted-leukocyte-reduced packed red blood cells (OR-LRpRBC), oxygen-depleted-leukocyte-reduced packed red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced packed red blood cells (OCR-LRpRBC), oxygen and carbon dioxide-depleted-leukocyte-reduced packed red blood cells containing platelets (OCR-LRpRBC+PLT), and combinations thereof. In other embodiments, the OR-preserved RBCs include oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT).

[0070] In one embodiment, the patient requiring frequent blood transfusions is a trauma patient. In another embodiment, the patient requiring frequent blood transfusions is a transplant patient. In yet another embodiment, the patient requiring frequent blood transfusions is a cardiac surgery patient. In yet another embodiment, the patient requiring frequent blood transfusions is an obstetric patient. In yet another embodiment, the patient requiring frequent blood transfusions is a gastrointestinal (GI) surgery patient. In yet another embodiment, the patient is an orthopedic surgery patient.

[0071] In one embodiment, the patient requiring frequent transfusions is a trauma patient. In another embodiment, the patient requiring frequent transfusions is a hemorrhagic trauma patient. In yet another embodiment, the patient requiring frequent transfusions is a blunt trauma patient.

[0072] In one embodiment, reducing cytokines in oxygen-depleted stored concentrated red blood cells provides an improvement in the treatment of cancer patients requiring blood transfusions. In the art, cytokines are known to be associated with negative patient outcomes in cancer patients receiving perioperative transfusions for surgical treatment. In one embodiment, a blood product with reduced oxygen and cytokines is provided to a cancer patient before surgery. In another embodiment, a blood product with reduced oxygen and cytokines is provided to a cancer patient during surgery. In yet another embodiment, a blood product with reduced oxygen and cytokines is provided to a cancer patient after surgery.

[0073] In embodiments of this disclosure, 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) have higher levels of 2,3-DPG compared to conventionally stored leukocyte-reduced 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 embodiment, 2,3-DPG levels are maintained at more than 50% of physiological levels for up to 4 weeks. In embodiments of this disclosure, improved 2,3-DPG levels are maintained for 2 weeks. In other embodiments, 2,3-DPG levels are maintained for 3 weeks. In one embodiment, the 2,3-DPG level in oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) is at least 80% of the 2,3-DPG level in blood on day zero. In another embodiment, the 2,3-DPG level in oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) is at least 5 to 20 DPG μmol / gHb.

[0074] Also provided and included in this disclosure are depleted whole blood (OR-WB) having lower levels of biological response modifiers (BRMs) compared to conventionally stored whole blood, including oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT). In certain embodiments, the BRM present in oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is about half the level of conventionally stored blood after 21 days. In one embodiment, 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) have relatively unchanged cytokine levels after 10 days of storage under anaerobic conditions. In another embodiment, cytokine levels remain relatively unchanged after 30 days of storage. In yet another embodiment, cytokine levels remain relatively unchanged after 40 days of storage. As used herein, “relatively unchanged” means that the cytokine concentration normalized to hemoglobin levels is within one standard deviation of the initial normalized concentration of the cytokine.

[0075] In certain embodiments, oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood with platelets (OCR-LRWB+PLT) have lower levels of the cytokine eotaxin compared to conventionally stored whole blood. In one embodiment, the level of eotaxin in oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood with platelets (OCR-LRWB+PLT) is about half the level of eotaxin present in conventionally stored blood after 21 days, normalized to hemoglobin concentration. In one embodiment, 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is approximately 25% or less of the level of eotaxin present in conventionally stored blood after 40 days.

[0076] In certain embodiments, oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood with platelets (OCR-LRWB+PLT) have lower levels of cytokines (RANTES, activated regulated normal T cell expression and secretions) compared to conventionally stored whole blood. In one embodiment, the levels of RANTES in oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood with platelets (OCR-LRWB+PLT) are about half the levels of RANTES in conventionally stored blood after 21 days, normalized to hemoglobin concentration. In one embodiment, the levels of rantine (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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are approximately 25% or less of the levels of rantine present in conventionally stored blood after 40 days.

[0077] In certain embodiments, oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have lower levels of monocyte chemotactic protein-1 (MCP-1) compared to conventionally stored whole blood. In one embodiment, the level of MCP-1 in oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is about half the level of MCP-1 present in conventionally stored blood after 21 days, normalized to hemoglobin concentration. In one embodiment, 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), or 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.

[0078] In certain embodiments, oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood with platelets (OCR-LRWB+PLT) have lower levels of angiogenin compared to conventionally stored whole blood. In one embodiment, the levels of angiogenin in oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood with platelets (OCR-LRWB+PLT) are about half the levels of angiogenin present in conventionally stored blood after 21 days, normalized to hemoglobin concentration. In one embodiment, the levels 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are approximately 25% or less of the levels of angiogenin present in conventionally stored blood after 40 days.

[0079] In certain embodiments, oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood with platelets (OCR-LRWB+PLT) have lower levels of tumor necrosis factor α (TNF-α) compared to conventionally stored whole blood. In one embodiment, the level of TNF-α in oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood with platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood with platelets (OCR-LRWB+PLT) is about half the level of TNF-α present in conventionally stored blood after 21 days, normalized to hemoglobin concentration. In one embodiment, 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is approximately 25% or less of the level of TNF-α present in conventionally stored blood after 40 days.

[0080] In certain embodiments, oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) have lower levels of epidermal growth factor (EGF) compared to conventionally stored whole blood. In one embodiment, the level of EGF in oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) is about half the level of EGF present in conventionally stored blood after 21 days, normalized to hemoglobin concentration. In one embodiment, 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is approximately 25% or less of the level of EGF present in conventionally stored blood after 40 days.

[0081] In certain embodiments, oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have lower levels of soluble CD40 ligand (sCD40L) compared to conventionally stored whole blood. In one embodiment, the level of sCD40L in oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is about half the level of sCD40L present in conventionally stored blood after 21 days, normalized to hemoglobin concentration. In one embodiment, 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is approximately 25% or less of the level of sCD40L present in conventionally stored blood after 40 days.

[0082] In certain embodiments, 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) have lower levels of platelet-derived growth factor (PDGF) compared to conventionally stored whole blood. In one embodiment, 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is about half the level of PDGF present in conventionally stored blood after 21 days, normalized to hemoglobin concentration. In one embodiment, the levels 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are approximately 25% or less of the levels of PDGF present in conventionally stored blood after 40 days.

[0083] This disclosure provides and includes oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB+PLT) containing platelets, oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood (OCR-LRWB+PLT) that, when transfused to a patient, provide a reduced inflammatory response compared to conventionally stored oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB+PLT), oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood (OCR-LRWB+PLT).

[0084] This disclosure provides and includes oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB) that provides a blood product having higher RBC deformability compared to conventionally stored blood products. In one particular embodiment, the blood product is a whole blood product. In another embodiment, the blood product is leukocyte-reduced whole blood. In yet another embodiment, the blood product is leukocyte-reduced whole blood and platelet-reduced whole blood. In a further embodiment, the blood product is leukocyte-reduced packed red blood cells or white blood cells and platelet-reduced packed red blood cells.

[0085] This disclosure provides and includes oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB) having coagulation parameters that, when measured by thromboelastography (TEG), are at least 75% of the coagulation parameters of conventionally stored whole blood. In one embodiment, the TEG coagulation parameters 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 equivalent to those of conventionally stored blood. In yet another embodiment, 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-extracted whole blood containing platelets (OCR-LRWB+PLT) have higher TEG coagulation parameters than conventionally stored blood. In one embodiment, the TEG angle is greater than 40 degrees. In another embodiment, the TEG kinetics (K) are less than 5 minutes. In another embodiment, the TEG K is between 1 and 5 minutes. In another embodiment, the TEG maximum amplitude (TEG MA) is greater than 50 mm. In another embodiment, the TEG maximum amplitude (TEG MA) is less than 70 mm. In another embodiment, the TEG maximum amplitude (TEG MA) is between 30 and 65 minutes. In another embodiment, the TEG reaction time (TEG R) is less than 10 minutes. In another embodiment, the TEG reaction time (TEG R) is less than 8 minutes. In yet another embodiment, the TEG reaction time (TEG R) is at least 3 minutes. In a further embodiment, the TEG reaction time (TEG R) is between 4 and 8 minutes.

[0086] This disclosure provides and includes oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB) having coagulation parameters that, when measured by prothrombin time (PT), are at least 75% of the coagulation parameters of conventionally stored whole blood. In one embodiment, 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), or 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 embodiment, 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) have a longer PT than conventionally stored blood. In yet another embodiment, oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB) has a PT of less than 15 seconds. In yet another embodiment, oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB) has a PT of more than 5 seconds. In yet another embodiment, oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB) has a PT of 10 to 15 seconds.

[0087] This disclosure provides and includes oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), which have coagulation parameters that, when measured by partial thromboplastin time (PTT), are at least 75% of those of conventionally stored whole blood. In one embodiment, the PTT of oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is equivalent to that of conventionally stored blood. In yet another embodiment, 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) have a longer PTT than conventionally stored blood. In yet another embodiment, 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) have a PTT longer than 25 seconds. In another embodiment, oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have a PTT of less than 40 seconds.In another embodiment, 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) have a PTT of 32-42 seconds.

[0088] This disclosure provides and includes oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), having whole blood (OR-WB) having coagulation parameters that, when measured by the level of fibrinogen activity, are at least 75% of the coagulation parameters of conventionally stored whole blood. In one embodiment, the fibrinogen activity of oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) is equivalent to that of conventionally stored blood. In yet another embodiment, 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) have a fibrinogen activity higher than that of conventionally stored blood. In yet another embodiment, 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) have a fibrinogen level of at least 200 mg / ml. In another embodiment, oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) have a fibrinogen level of at most 400 mg / ml.In another embodiment, 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) have fibrinogen levels of 250-350 mg / ml. In another embodiment, 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) have fibrinogen levels of 250-300 mg / ml.

[0089] This disclosure provides and includes oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), which have coagulation parameters that, when measured by D-dimer analysis, are at least 75% of those of conventionally stored whole blood (OR-WB). In one embodiment, the D-dimer values ​​of oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are equivalent to those of conventionally stored blood. In yet another embodiment, 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) have higher D-dimer values ​​than conventionally stored blood.

[0090] This 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), which have coagulation parameters that, when measured by a thrombin generation test, are at least 75% of the coagulation parameters of conventionally stored whole blood (OR-WB). In one embodiment, the thrombin generation 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 equivalent to that of conventionally stored blood. In yet another embodiment, 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) have higher thrombin production values ​​than conventionally stored blood.

[0091] This disclosure provides and includes oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), which, when measured by a platelet agglutinator, have platelet function parameters that are at least 75% of the platelet function parameters of conventionally stored whole blood. In one embodiment, the platelet function parameters of oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are equivalent to the platelet function parameters of conventionally stored blood. In yet another embodiment, 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) have higher platelet function parameters than conventionally stored blood.

[0092] This 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), having coagulation factor levels that are at least 75% of the levels of coagulation factors in conventionally stored blood. In one embodiment, the coagulation factor levels are equivalent to those of coagulation factors in conventionally stored blood. In yet another embodiment, 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) have platelet function parameters higher than those of conventionally stored blood. Although not constrained by theory, it is thought that the oxidative degradation of coagulation factors is prevented or reduced in oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT), providing higher levels of coagulation factor activity. Methods for evaluating the effects of treatment on coagulation ability are known in the art and are described, for example, in 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).

[0093] This 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), having a level of factor V with specific activity that is at least 75% of the level of factor V activity present in conventionally stored blood. In one embodiment, the specific activity of factor V is equivalent to that of conventionally stored blood. In further embodiments, 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) have higher specific activity of factor V than conventionally stored blood platelet function parameters. Methods for measuring the specific activity of factor V are known in the art.

[0094] This 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), having a level of factor V with specific activity at least 75% of the level of factor VIII activity present in conventionally stored blood. In one embodiment, the specific activity of factor VIII is equivalent to that of conventionally stored blood. In another embodiment, 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) have higher specific activity of factor VIII than conventionally stored blood platelet function parameters. Methods for measuring the specific activity of factor VIII are known in the art. In one embodiment, 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) have a specific activity of factor V of less than 40% after 21 days of storage.

[0095] This disclosure provides and includes oxygen-depleted-leukocyte-reduced whole blood (OR-LRWB), oxygen-depleted-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-depleted-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), having a level of antithrombin (AT) with specific activity that is at least 75% of the level of AT activity present in conventionally stored blood. In one embodiment, the specific activity of AT is equivalent to that of conventionally stored blood. In further embodiments, 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) have higher specific activity of AT than conventionally stored blood platelet function parameters. Methods for measuring the specific activity of AT are known in the art.

[0096] This disclosure provides and includes oxygen-decreased-leukocyte-reduced whole blood (OR-LRWB), oxygen-decreased-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-decreased-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-decreased-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), having a level of factor XIV (autoprothrombin IIA or protein C) with specific activity at least 75% of the level of factor XIV activity present in conventionally stored blood. In one embodiment, the specific activity of factor XIV is equivalent to that of conventionally stored blood. In yet another embodiment, 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) have higher specific activity of factor XIV than conventionally stored blood platelet function parameters. Methods for measuring the specific activity of factor XIV are known in the art.

[0097] This disclosure provides and includes oxygen-decreased-leukocyte-reduced whole blood (OR-LRWB), oxygen-decreased-leukocyte-reduced whole blood containing platelets (OR-LRWB+PLT), oxygen and carbon dioxide-decreased-leukocyte-reduced whole blood (OCR-LRWB), or oxygen and carbon dioxide-decreased-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT), having a level of von Willebrand factor (vWF) with specific activity at least 75% of the level of vWF activity present in conventionally stored blood. In one embodiment, the specific activity of vWF is equivalent to that of conventionally stored blood. In yet another embodiment, 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) have a specific activity of vWF higher than the platelet function parameters of conventionally stored blood. In yet another embodiment, 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) have a specific activity of vWF lower than the platelet function parameters of conventionally stored blood. Methods for measuring the specific activity of vWF are known in the art.

[0098] This disclosure provides and includes methods for extending the shelf life of whole blood from the current two weeks to three weeks or more. The oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) of this disclosure provide patient outcomes at three weeks that are equivalent to those provided by whole blood stored for two weeks under conventional conditions.

[0099] As provided herein, oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) have fewer side effects on transfusion recipients compared to conventionally stored blood. In one embodiment, oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) show less inflammatory response after 2 weeks of storage compared to conventionally stored blood. In another embodiment, the inflammatory response is reduced after 3 weeks compared to conventionally stored blood. In one embodiment, 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) can be stored for longer than 3 weeks and, after 2 weeks, retain levels of inflammatory response compared to conventionally stored blood.

[0100] As provided herein, 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) have fewer side effects on transfusion recipients compared to conventionally stored blood. In one embodiment, 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) exhibit lower immunomodulatory effects after 2 weeks of storage compared to conventionally stored blood. In another embodiment, immunomodulatory effects are lower after 3 weeks of storage compared to conventionally stored blood. In one embodiment, 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) can be stored for longer than 3 weeks and, after 2 weeks, retain levels of immunomodulatory activity compared to conventionally stored blood.

[0101] The methods and whole blood products of this disclosure provide improved patient outcomes when transfused. In particular, oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) provide improved survival in cancer patients when provided in perioperative transfusions. In certain embodiments, oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) provide reduced mortality and improved survival when provided perioperatively to pancreatic cancer patients. While not constrained by theory, the reduction in mortality is a result of a combination of decreased cytokine levels and improved oxygen transport and delivery due to increased 2,3-DPG and ATP levels.

[0102] In one embodiment, blood for transfusion to cancer patients requiring transfusion has low levels of activated regulated normal T cell expression and secreted (RANTES) cytokines. In one embodiment, the RANTES level is equivalent to the RANTES level present at the start of storage. In another embodiment, the RANTES levels in oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) are lower than the RANTES levels present in conventionally stored blood. In one embodiment, the RANTES level remains lower throughout the storage period than the RANTES level present in conventionally stored blood. In another embodiment, RANTES does not increase during storage.

[0103] In one embodiment, blood for transfusion to cancer patients requiring transfusion has low levels of CC chemokines, namely eotaxin, an eosinophil chemotactic protein. In one embodiment, low levels of eotaxin in oxygen-reduced-leukocyte-reduced whole blood (OR-LRWB), oxygen-reduced-leukocyte-reduced whole blood with 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 with platelets (OCR-LRWB+PLT) are eotaxin-1, also known as CC motif chemokine 11. In one embodiment, the eotaxin level is equivalent to the level of eotaxin present at the start of storage. In another embodiment, the eotaxin levels 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT) are lower than the levels of eotaxin present in conventionally stored blood. In one embodiment, the eotaxin levels remain lower throughout the storage period than the levels of eotaxin present in conventionally stored blood. In another embodiment, eotaxin does not increase during the storage period.

[0104] The methods and whole blood products described herein, when transfused, provide relief from multi-organ dysfunction syndrome and improve patient outcomes. 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), or oxygen and carbon dioxide-reduced-leukopenic whole blood containing platelets (OCR-LRWB+PLT) provide relief from multi-organ dysfunction syndrome in trauma patients when provided in perioperative transfusions. In certain embodiments, 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), when provided during emergency treatment, provide relief of multi-organ dysfunction syndrome in trauma patients.

[0105] This disclosure provides and includes a method for preparing oxygen-depleted, leukocyte-depleted whole blood, comprising obtaining a unit of whole blood containing an anticoagulant, filtering the whole blood to produce leukocyte-depleted whole blood, depleting oxygen from the leukocyte-depleted whole blood, and storing the oxygen-depleted, leukocyte-depleted whole blood under anaerobic conditions.

[0106] This disclosure provides and includes a method for preparing oxygen-reduced, leukocyte-reduced whole blood having a pre-storage oxygen saturation (SO2) of 30% or less. Whole blood obtained from a donor by venipuncture has an oxygen saturation (SO2) in the range of approximately 30% to approximately 70% saturated oxygen. In certain embodiments, the SO2 is reduced to 25% or less. In certain embodiments, the SO2 is reduced to 20% or less. In certain embodiments, the SO2 is reduced to 15% or less. In other embodiments, the SO2 is reduced to 10% or less. In yet another embodiment, the SO2 is reduced to 5% or less.

[0107] Also provided and included in this disclosure are oxygen-reduced and carbon dioxide-reduced-leukocyte-reduced whole blood compositions and methods for preparing such compositions. In certain embodiments, the SO2 value is less than 20% and the carbon dioxide partial pressure is less than 60 mmHg. In other embodiments, the carbon dioxide partial pressure is between 10 and 60 mmHg. In yet another embodiment, the carbon dioxide partial pressure is between 20 and 40 mmHg. Also included are whole blood compositions and methods that provide SO2 of 15% or less and a carbon dioxide partial pressure of 10 to 60 mmHg. In yet another embodiment, the method and composition comprises a whole blood product having SO2 of 15% or less and a carbon dioxide partial pressure of 20 to 40 mmHg. In yet another embodiment, the blood compositions and methods of this disclosure have SO2 of 10% or less and a carbon dioxide partial pressure of 10 to 60 mmHg. In yet another embodiment, the blood compositions and methods of this disclosure have SO2 of 10% or less and a carbon dioxide partial pressure of 20 to 40 mmHg. In a further embodiment, the blood composition and method of the present disclosure have a SO2 content of 5% or less and a carbon dioxide partial pressure of 10 to 60 mmHg. In another embodiment, the blood composition and method of the present disclosure have a SO2 content of 5% or less and a carbon dioxide partial pressure of 20 to 40 mmHg.

[0108] Furthermore, oxygen-reduced and carbon dioxide-reduced - leukocyte-reduced whole blood compositions and methods for preparing these compositions are also provided and included in this disclosure. In certain embodiments, the SO2 value is ≤20% and the carbon dioxide partial pressure is 1–60 mmHg. In other embodiments, the carbon dioxide partial pressure is 10–60 mmHg. In yet another embodiment, the carbon dioxide partial pressure is 20–40 mmHg or 1–20 mmHg. Also included are whole blood compositions and methods that provide SO2 ≤15% and a carbon dioxide partial pressure of 10–60 mmHg. In certain embodiments, the SO2 value is ≤15% and the carbon dioxide partial pressure is 1–60 mmHg. In yet another embodiment, the method and composition include a whole blood product having SO2 ≤15% and a carbon dioxide partial pressure of 20–40 mmHg or 1–20 mmHg. In yet another embodiment, the blood compositions and methods of this disclosure have SO2 ≤10% and a carbon dioxide partial pressure of 1–60 mmHg or 10–60 mmHg. In other embodiments, the blood composition and method of this disclosure has less than 10% SO2 and a partial pressure of carbon dioxide of 20-40 mmHg or 1-20 mmHg. In further embodiments, the blood composition and method of this disclosure has less than 5% SO2 and a partial pressure of carbon dioxide of 1-60 mmHg or 10-60 mmHg. In other embodiments, the blood composition and method of this disclosure has less than 5% SO2 and a partial pressure of carbon dioxide of 20-40 mmHg or 1-20 mmHg.

[0109] In particular, and as is evident in Figures 2A, 2B, 3A, and 3B, the ATP level in the stored oxygen-reduced blood depends on the CO2 partial pressure. Specifically, oxygen depletion to about 10% SO2 and carbon dioxide depletion to about 25 mmHg result in an increase in 2,3-DPG levels that persist beyond 21 days, while ATP is reduced to about half of its initial level. See Figures 2G and 3D. Therefore, this disclosure provides and includes oxygen depletion to about 5% SO2 level and carbon dioxide depletion to about 30-40 mmHg partial pressure to obtain oxygen-reduced whole blood that has elevated levels of 2,3-DPG and retains at least 50% of the initial ATP concentration up to day 20. In other embodiments, the CO2 partial pressure can be adjusted to maintain an ATP level that is at least 75% of the initial ATP value. The adjustment of the CO2 level can be determined experimentally by those skilled in the art, taking this disclosure into consideration.

[0110] Long-term cryogenic storage under conventional conditions is known to impair the deformability of stored red blood cell block (RBCs), potentially hindering their ability to perfuse the microvascular network and deliver oxygen to tissues and vital organs during transfusion. Oxidative damage is considered a major cause of RBC biomechanical loss; therefore, storing RBCs under oxygen-reduced (OR) and oxygen- and carbon dioxide-reduced (OCR) conditions mitigates oxidative damage, thereby preserving superior intrinsic rheological properties compared to conventional (aerobic) storage. For this study, we used an extracorporeal microfluidic system that replicates the capillary bed of in vivo microvessels to demonstrate the effects of oxygen reduction on stored cells.

[0111] This disclosure provides and includes a method for managing a blood bank that improves the availability of blood products for trauma patients and patients requiring frequent transfusions, thereby providing overall blood resource conservation. Component blood products can be prepared from stored whole blood according to this application and can be used for transfusion or incorporated into a large transfusion kit. In addition to improvements in blood chemistry (e.g., lower hemolysis, improved 2,3-DPG), the method provides improved homeostasis and enhanced deformability.

[0112] In embodiments thereof, the method provides for maintaining a stock of blood units containing oxygen-depleted whole blood and an anticoagulant as described above, providing one or more blood units from the stock for the treatment of a patient, and reusing blood units from the stock to prepare a component-separated oxygen-depleted blood unit containing oxygen-depleted plasma and oxygen-depleted leukocyte-depleted packed red blood cells (OR-LRpRBC+PLT) containing platelets. In embodiments, the anticoagulant includes citrate-phosphate-dextrose (CPD), citrate-phosphate-dextrose containing adenine (CPDA-1), or CP2D.

[0113] In one embodiment, the Specified provides a method for maintaining a stock of blood units containing oxygen- and carbon dioxide-reduced-leukocyte-reduced whole blood and an anticoagulant as described above, providing one or more blood units from the stock for the treatment of a patient, and reusing blood units from the stock to prepare a component-separated oxygen-reduced blood unit containing oxygen- and carbon dioxide-reduced plasma and oxygen- and carbon dioxide-reduced-leukocyte-reduced packed red blood cells (OCR-LRpRBC+PLT) containing platelets. In one embodiment, the anticoagulant includes citrate-phosphate-dextrose (CPD), citrate-phosphate-dextrose-adenine (CPDA-1) containing adenine, or the anticoagulant citrate-phosphate-2-dextrose (CP2D).

[0114] This specification further provides for the preparation of one or more large-volume transfusion kits, as described later, comprising oxygen-depleted plasma, oxygen-depleted-leukocyte-depleted packed red blood cells containing platelets (OR-LRpRBC+PLT), oxygen and carbon dioxide-depleted plasma, and oxygen and carbon dioxide-depleted-leukocyte-depleted packed red blood cells containing platelets (OCR-LRpRBC+PLT).

[0115] In an embodiment of the present invention, unused blood in stock is reused after a certain period of time. In one particular embodiment, if the anticoagulant is CPD, the blood unit is reused before it has been stored for three weeks. In another embodiment, if the anticoagulant is CPDA-1, the blood unit is reused before it has been stored for five weeks. In yet another embodiment, the blood unit is reused after two weeks or earlier. In one embodiment, the reuse of the blood unit takes place between two days and one week. In another embodiment, the reuse takes place between two days and two weeks. In several embodiments, the reuse takes place between one and two weeks. The timing of reuse may vary in accordance with the turnover rate and needs of the blood facility.

[0116] The reuse process is preferably carried out under anaerobic conditions, although it may also be carried out under aerobic conditions. Aerobic conditions may offer cost savings, but at the same time, they may be suitable for facilities with higher turnover rates. In facilities with high turnover rates, the recovered blood components may be used immediately after the reuse process, so further storage of blood under anaerobic conditions offers little additional benefit.

[0117] Methods for managing blood banks further provide for the preparation of mass transfusion kits, which will be detailed later.

[0118] This disclosure provides and includes a method for supplying blood products for transfusion medicine, comprising: preparing oxygen-depleted leukocyte-reduced whole blood (OR-LRWB+PLT) by depleting oxygen from leukocyte-reduced whole blood; storing the oxygen-depleted leukocyte-reduced whole blood (OR-LRWB+PLT) for a certain period of time; and providing the stored blood to patients who need it. In a particular embodiment, the leukocyte reduction step includes platelet reduction to produce oxygen-depleted leukocyte-reduced whole blood (OR-LRWB).

[0119] This disclosure provides and includes a method for supplying blood products for transfusion medicine, comprising: preparing oxygen- and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB+PLT) by depleting oxygen and carbon dioxide from leukocyte-reduced whole blood; storing the oxygen- and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB+PLT) for a certain period of time; and providing the stored blood to patients in need. In a particular embodiment, the leukocyte-reducing step includes platelet reduction to produce oxygen- and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB).

[0120] This disclosure provides and includes a method for supplying blood products for transfusion medicine, comprising: preparing oxygen-depleted leukocyte-depleted whole blood (OR-LRWB+PLT) by depleting oxygen from leukocyte-depleted whole blood; storing the oxygen-depleted leukocyte-depleted whole blood (OR-LRWB+PLT) for a certain period of time; and preparing oxygen-depleted leukocyte-depleted packed red blood cells (OR-LRpRBC+PLT) containing platelets. In a particular embodiment, the leukocyte-depletion step includes platelet reduction to produce oxygen-depleted leukocyte-depleted packed red blood cells (OR-LRpRBC).

[0121] This disclosure provides and includes a method for supplying blood products for transfusion medicine, comprising: preparing oxygen- and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB+PLT) by depleting oxygen and carbon dioxide from leukocyte-reduced whole blood; storing oxygen- and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB+PLT) for a certain period of time; and preparing oxygen- and carbon dioxide-reduced leukocyte-reduced packed red blood cells (OCR-LRpRBC+PLT) containing platelets. In a particular embodiment, the leukocyte-reducing step includes platelet reduction to produce oxygen- and carbon dioxide-reduced leukocyte-reduced packed red blood cells (OCR-LRpRBC).

[0122] Where provided herein, OR-LRpRBC+PLT, OR-LRpRBC, OCR-LRpRBC+PLT, and OCR-LRpRBC may be returned to the blood product storage for supply and stored for a certain period until required for use by a patient. In embodiments of this disclosure, the total storage time as a whole blood product or as a concentrated RBC product may be up to 6 weeks. In some embodiments, the second storage period is 2 to 4 weeks.

[0123] A method for supplying blood products includes and provides depleting either oxygen or oxygen and carbon dioxide. The oxygen level for a method for supplying blood products is described in detail above. In one particular embodiment, the SO2 level is reduced to 20% or less and the carbon dioxide partial pressure is less than 60 mmHg. In another embodiment, the carbon dioxide partial pressure is 10 to 60 mmHg. In yet another embodiment, the carbon dioxide partial pressure is 20 to 40 mmHg. Also included is a method that provides SO2 of 15% or less and a carbon dioxide partial pressure of 10 to 60 mmHg. In yet another embodiment, the method of this disclosure provides a blood product having SO2 of 15% or less and a carbon dioxide partial pressure of 20 to 40 mmHg. In yet another embodiment, the method of this disclosure provides a blood product having SO2 of 10% or less and a carbon dioxide partial pressure of 10 to 60 mmHg. In yet another embodiment, a method for supplying blood products provides SO2 of 10% or less and a carbon dioxide partial pressure of 20 to 40 mmHg. In a further embodiment, the method provides a partial pressure of 5% or less SO2 and 10 to 60 mmHg of carbon dioxide. In another embodiment, the method provides a partial pressure of 5% or less SO2 and 20 to 40 mmHg of carbon dioxide.

[0124] This disclosure provides and includes novel blood compositions obtained during the blood component recovery process of OR-LRWB+PLT and OCR-LRWB+PLT. As provided above, conventional whole blood products have an FDA-approved shelf life (3 weeks for WB in CPD and 5 weeks in CPDA1), but physicians using WB limit its shelf life to 2 to 14 days. In conventional storage, blood is often discarded. This disclosure shows that OR-LRWB+PLT and OCR-LRWB+PLT can be processed using conventional component separation methods that have been improved to keep the blood in an OR or OCR depleted state. Generally, the methods are improved to incorporate oxygen-impermeable and oxygen and carbon dioxide-impermeable barriers to the components and features to prevent oxygen intrusion. Appropriate methods can be found, for example, in International Patent Application No. PCT / US2016 / 021794, filed on 10 March 2016, and International Patent Application No. PCT / US2016 / 029069, filed on 22 April 2016, both of which are incorporated herein by reference in their entirety.

[0125] In a manner consistent with this disclosure, 1 × 10 5 A blood composition is provided that includes oxygen-depleted packed red blood cells and platelets with a leukocyte count of less than 1 / L. Such compositions are obtained from OR-LRWB+PLT and OCR-LRWB+PLT. In one embodiment, the leukocyte level is 1 × 10⁻⁶. 4 The oxygen saturation is less than / L leukocytes. In an aspect of this disclosure, the oxygen saturation of oxygen-depleted leukocyte-depleted packed erythrocytes (OR-LRpRBC+PLT) containing platelets is less than 30%. In one aspect, the oxygen saturation of oxygen-depleted leukocyte-depleted packed erythrocytes (OR-LRpRBC+PLT) containing platelets is less than 20%. In one aspect, the oxygen saturation of oxygen-depleted leukocyte-depleted packed erythrocytes (OR-LRpRBC+PLT) containing platelets is less than 10%. In a further aspect, the oxygen saturation of oxygen-depleted leukocyte-depleted packed erythrocytes (OR-LRpRBC+PLT) containing platelets is less than 5%.

[0126] This disclosure provides and includes oxygen and carbon dioxide reduced-leukocyte reduced packed red blood cells (OCR-LRpRBC+PLT) containing platelets having less than 30% SO2 and a partial pressure of carbon dioxide at storage of less than 60 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 30% and a partial pressure of carbon dioxide at storage of 20-40 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 30% and a partial pressure of carbon dioxide at storage of 0-20 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 20% and a partial pressure of carbon dioxide at storage of less than 60 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 20% and a partial pressure of carbon dioxide at storage of 20-40 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 20% and a partial pressure of carbon dioxide at storage of 0-20 mmHg. In another embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 15% and a storage partial pressure of carbon dioxide of less than 60 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 15% and a storage partial pressure of carbon dioxide of 20-40 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 15% and a storage partial pressure of carbon dioxide of 0-20 mmHg. In another embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 10% and a storage partial pressure of carbon dioxide of less than 60 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 10% and a storage partial pressure of carbon dioxide of 20-40 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 10% and a storage partial pressure of carbon dioxide of 0-20 mmHg. In another embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 5% and a storage partial pressure of carbon dioxide of less than 60 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 5% and a storage partial pressure of carbon dioxide of 20-40 mmHg. In one embodiment, OCR-LRpRBC+PLT has an oxygen saturation of less than 5% and a storage partial pressure of carbon dioxide of 0-20 mmHg.

[0127] Platelet-containing oxygen-reduced leukocyte-reduced packed red blood (OR-LRpRBC+PLT) and platelet-containing oxygen and carbon dioxide-reduced leukocyte-reduced whole blood (OCR-LRWB+PLT) typically further contain additives. Suitable additives according to this disclosure include AS-1, AS-3 (Nutricel®), AS-5, SAGM, PAGG-SM, PAGG-GM, MAP, AS-7, ESOL-5, EAS61, OFAS1, OFAS3, and combinations thereof. In one embodiment, the additive is added at the time of component separation. In one embodiment, the additive is AS-1. In another embodiment, the additive is AS-3. In yet another embodiment, the additive is SAGM.

[0128] The methods and compositions of this disclosure provide and include the preparation of “mass transfusion kits” (MTKs) having improved properties compared to kits prepared from conventional compositions. The mass transfusion kits of this disclosure can be prepared in various configurations depending on clinical needs. The MTKs of this disclosure are stored under anaerobic or anaerobic and carbon dioxide-free conditions until prepared for use. OR and OCR conditions can be maintained by sealing in an impermeable enclosure with or without appropriate absorbent material. The MTKs of this disclosure may be re-oxygenated before use or may be used directly. Generally, this specification provides mass transfusion kits optimized to deliver RBCs having improved 2,3-DPG levels. Such kits are prepared from component blood products obtained from oxygen- and carbon dioxide-reduced-leukocyte-reduced whole blood (OCR-LRWB+PLT) containing platelets. Alternatively, to produce kits with higher levels of ATP, kits may be prepared from component blood products obtained from oxygen- and carbon dioxide-reduced-leukocyte-reduced whole blood (OR-LRWB+PLT) containing platelets. Kits prepared using the methods described herein provide homeostatic platelets along with oxygen-depleted preserved red blood cells. Thus, the mass transfusion kits described herein can increase platelet availability without additional dilution, while further providing higher quality RBCs (e.g., more deformable, more 2,3-DPG, less storage impairment). Importantly, the recovery of blood components from oxygen-depleted whole blood according to this disclosure increases the availability of transfusion products for trauma patients, conserving and preserving precious, limited resources. As previously noted, conventional mass transfusion kits contain a certain volume of plasma, a certain volume of pRBCs, and a certain volume of platelets in a 1:1:1 ratio, with the amounts of the three components corresponding to units of “reconstituted blood” when transfused sequentially or in parallel to patients in need. Reconstituted blood does not directly correspond to whole blood with a higher level of anticoagulant and without additives. Furthermore, reconstituted blood typically contains a larger volume than typical whole blood units.The reconstituted blood of this disclosure is an improvement over conventional reconstituted blood because it provides additional platelets in the pRBC fraction (e.g., oxygen- and carbon dioxide-reduced leukocyte-reduced packed red blood cells containing platelets (OCR-LRpRBC+PLT) and oxygen- and carbon dioxide-reduced leukocyte-reduced packed red blood cells containing platelets (OR-LRpRBC+PLT)). Such cryopreserved platelets undergo a change commonly known as platelet preservation disorder (PSL), and cryopreserved platelets are rapidly reduced from circulation in the body. Importantly, cryopreserved platelets have been reported to retain their ability to aggregate and to have resistance to increased aggregation and deaggregation. Therefore, the blood components obtained from oxygen-reduced whole blood of this specification, either alone or in combination with conventional platelets, provide further advantages during transfusion for trauma.

[0129] This disclosure provides and includes a massive transfusion kit comprising a certain volume of oxygen-depleted, leukocyte-depleted packed red blood (OR-LRpRBC+PLT) containing platelets or oxygen and carbon dioxide-depleted, leukocyte-depleted whole blood (OCR-LRWB+PLT) containing platelets or a combination thereof. In one embodiment, the massive transfusion kit provides a certain volume of plasma and a certain volume of LRpRBC+PLT. In one embodiment, the volume of plasma and the volume of LRpRBC+PLT are in a 1:1 ratio. In another embodiment, the volume ratio of plasma to LRpRBC+PLT is 1:1 to 1:2. In one embodiment, the volume ratio of plasma to LRpRBC+PLT is 1:2.

[0130] This disclosure provides and includes a massive transfusion kit containing additional platelets, along with plasma and oxygen-depleted, leukocyte-depleted packed red blood cells containing platelets (OR-LRpRBC+PLT) or oxygen and carbon dioxide-depleted, leukocyte-depleted whole blood containing platelets (OCR-LRWB+PLT).

[0131] The mass transfusion kit of this disclosure provides a certain volume of plasma. The plasma for MTK may be either fresh plasma or thawed fresh frozen plasma (FFP). This specification provides for obtaining plasma for MTK from either a conventional source (e.g., non-oxygen-reduced) or an oxygen-reduced or oxygen and carbon dioxide-reduced source. In one embodiment, the plasma for MTK of this disclosure may 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), or oxygen and carbon dioxide-reduced-leukocyte-reduced whole blood containing platelets (OCR-LRWB+PLT). Although not constrained by theory, plasma obtained from an oxygen-reduced source has lower levels of preservation impairment, including, for example, lower levels of cytokines, isoprostanes, and microparticles. Where provided herein, MTK having plasma, platelets, and pRBCs is provided in a volume ratio of 1:1:1 or 1:1:2. Those skilled in the art will understand that the MTK of this disclosure, like conventional MTK, is designed to provide an equivalent of one unit of blood. It should be noted that any total volume may be selected while maintaining the described ratios necessary to be equivalent to reconstituted blood. [Examples]

[0132] Example 1: Accumulation of cytokines, cell-free hemoglobin, and isoprostane in packed red blood cells during anaerobic storage. Fifteen pRBC units are collected from a healthy donor. Each unit is divided and stored as follows: one is stored under standard blood bank conditions (control), and the other is stored anaerobically according to the method described in Yoshida et al., “Anaerobic Storage of Red Blood Cells in a Novel Additive Solution Improves In vivo Recovery,” Transfusion 49:458-64 (2008) (test). Samples are reduced from the PRBC units using a sterile connection device at weeks 0, 1, 2, 3, and 6. Plasma samples are frozen for the following assays: single-batch assay for 22 cytokines using the Procarta Immunoassay Magnetic Bead kit, and 8-isoprostane F by mass spectrometry assay. 2α , and cell-free hemoglobin measured using HemoCue plasma / photometer (HemoCue AB, Angelholm, Sweden).

[0133] As shown in Figure 1A, eotaxin reached statistical significance at week 2 (86.6 pg / ml - control (c), 64.9 - study (t), p-value -0.00213, statistical significance of p<0.05; day 42 (292-c, 112-t; p=0.000). As shown in Figure 1B, RANTES differed at all time points starting from day 3 (374.6-c, 55.1-t), with p-=0.00000, and a very large difference was observed on day 42 (3371). 6-c, 88.4-t; p<0.002). As shown in Figure 1C, the difference in cell-free hemoglobin is observed at week 2 (96.0 mg / dl-c, 41.7-t), p-=0.00001; day 42 (170-c, 63-t, p=0.0002). As shown in Figure 1D, the difference in isoprostane is shown on day 3 of storage (45.5 pg / ml-c, 32.1-t), p=0.00689; day 42 (101.9-c, 64.7-t, p=0.0048).

[0134] Example 2: Whole blood collection, leukocyte reduction, and gas depletion Collect one unit of blood from the donor patient in an anticoagulant solution containing either CPDA1 or CPDA, following a standard protocol that includes heparin tubule retrieval. Within four hours of the initial blood collection, reduce the leukocyte count from the collected blood containing the anticoagulant according to the manufacturer's instructions. Baseline ABL90 blood gases and metabolic parameters are determined from the donor's heparin tubule and whole blood product following a standard procedure. Refer to the BSL Handbook Procedure BSL-P024:Procedure Manual and the Radiometer ABL90 FLEX Gas Analyzer instructions.

[0135] Prepare anaerobic controls from each unit of leukopenic blood by transferring 120 ml of LRWB / CPDA-1 or LRWB / CPD to a 150 mL transport bag, which has been placed at room temperature (15°C to 30°C), and labeling as necessary.

[0136] The remaining LRWB LRWB / CPDA-1 or LRWB / CPD is transferred to a blood processing bag connected to the Sorin D100 and processed for oxygen depletion or oxygen and carbon dioxide depletion, and then processed at a flow rate of 700 ml / min for 5 minutes without gas to prepare a BOF-treated control. The resulting 120 g of BOF-treated blood is transferred to a 300 ml transport bag labeled as a BOF-treated control, which has been stored under anaerobic conditions. The remaining LRWB / CPDA-1 or LRWB / CPD is processed on the Sorin D100 at a peak flow rate of 700 ml / min with a gas flow rate of 3 L / min using a gas composition containing 5% CO2 / 95% N2 until the blood reaches approximately 5% SO2, and blood gas values ​​are measured at 3-5 minute intervals using a radiometer ABL90 FLEX Gas Analyzer. To reduce carbon dioxide levels, switch the gas mixture to 100% N2 for 1–4 minutes until SO2 reaches 5±1% and pCO2 reaches 30±3 mmHg, and monitor blood gas values ​​every 15–30 seconds to monitor the deoxygenation rate. Transfer the resulting 120 g of oxygen- and carbon dioxide-reduced LRWB / CPDA-1 or LRWB / CPD to a 300 ml transport bag that was previously stored under anaerobic conditions as described above, and label it ("C"). Further processing of LRWB / CPDA-1 or LRWB / CPD is performed on a Sorin D100 at a flow rate of 700 ml / min using 99% N2 and 1% O2 until LRWB / CPDA-1 or LRWB / CPD reaches 5±1% SO2 and pCO2 reaches 7±3 mmHg. The resulting 120g of oxygen- and carbon dioxide-reduced LRWB / CPDA-1 or LRWB / CPD is transferred to a 300ml transport bag that was previously stored under anaerobic conditions as described above, and labeled ("D"). Further samples are processed as described above using new Sorin D100. Immediately after preparing each sample, ABL90 blood gas levels are determined according to the manufacturer's instructions, and baseline SO2 and pCO2 levels (e.g., T0) are defined. Refer to the BSL Handbook Procedures. Samples for cytokine analysis are collected and stored at -80°C for later analysis.

[0137] All samples are analyzed as described later in Example 6.

[0138] Example 3: Storage of anaerobic test preparations Place the oxygen-reduced and oxygen- and carbon dioxide-reduced blood, which are in transport bags, in mesh, secure with elastic cords, and place them in an anaerobic canister along with four absorbent bags (Mitsubishi, SS-300). Seal the canister and remove air from it using an Alicat Gas Processing System. Refer to BSL Handbook Procedure BSL-P040: Procedure for Placing Blood Products in Anaerobic Storage in Canisters. Place the anaerobic and aerobic blood in a blood bank refrigerator at 1-6°C. Monitor the canister gauges daily to ensure they display 5±1 psi. Canisters below 2 psi should be adjusted to the standard procedure. Refer to BSL Handbook Procedure BSL-P040: Procedure for Placing Blood Products in Anaerobic Storage in Canisters.

[0139] Example 4: Sample Testing Test the samples at the specified time points: day 0 (T0), day 1, week 1, week 2, and week 3 after processing. Samples may be tested fresh or frozen for later testing, as required for the given tests. Tests include whole blood cell count (CBC) and thromboelastography (TEG).

[0140] Prepare platelet-rich plasma (PRP) for platelet aggregation immediately, following the manufacturer's instructions.

[0141] Perform coagulation screening and additional assays according to the manufacturer's instructions.

[0142] Prepare the cytokine sample immediately, following the manufacturer's instructions.

[0143] Example 5: ATP sample collection and measurement For ATP measurement, the sample is treated by deproteinization and precipitation. Precipitate 1 ml of the sample (e.g., LRWB / CPD or LRWB / CPDA-1 or the aforementioned sample) with 1.0 ml of ice-cold trichloroacetic acid (TCA) (12% w / v), vortex for 15-30 seconds, and incubate on ice for 5 minutes. Centrifuge the test tube containing the TCA / sample mixture at 3600 g for 5 minutes at 4°C. Process the sample immediately to minimize exposure to TCA. Transfer the clear supernatant to a pre-cooled test tube, flash-freeze in a dry ice alcohol bath, and store at -70°C.

[0144] Example 6: Improvement of deformability in stored RBCs preserved under oxygen-reduced conditions Nine individual whole blood units are obtained from healthy, consenting volunteers through standard 500 mL blood donations. The donated whole blood is processed into leukocyte-reduced red blood cell (LR-RBC) units according to standard AABB / FDA guidelines, and the resulting units are then divided into two equal parts. One half of each unit is deoxygenated with reduced O2 and CO2 as described in Examples 2 and 3.

[0145] The obtained samples are stored anaerobically at low temperatures, and the second half is stored aerobically at low temperatures as before. A pair of RBC units are stored in the blood bank refrigerator and evaluated weekly throughout the entire 6-week storage period. Before the test, the hematocrit value of all RBC samples was adjusted to 40% using physiological saline (0.9% NaCl; RBC-S). The deformability of RBC-S at the start of the test and during the test is determined as described in international patent application WO2013 / 177339, published on November 28, 2013. A high-speed image sequence (approximately 150 FPS) of the blood sample traversing the artificial microvascular network (AMVN) chip is recorded. The occlusion time, which is the time it takes for blood to flow through the vascular network obstructed by non-deformable cells, and the frequency of interruptions in blood perfusion through the vascular network (occlusion frequency) are determined.

[0146] The overall mass perfusion rate through the AMVN system was consistently faster with O2 and CO2-controlled blood compared to aerobically stored units, and the total occlusion time was consistently shorter with oxygen-reduced RBCs (Table 1). These results suggest that reduced oxygen levels in LR-RBC units mitigate the degradation of the biomechanical properties of erythrocytes during cryogenic storage.

[0147] The oxygen depletion and preservation process significantly slows the rate at which the rheological properties of RBCs deteriorate during cryogenic storage, making it possible to preserve the physiologically more appropriate biomechanical properties of red blood cells during storage. The improved deformability of RBCs, coupled with the benefits of whole blood transfusion, means that the function of preserved RBCs improves post-transfusion RBC retention and increases the ability of transfused RBCs to perfuse the microvascular system. Table 1: Perfusion rate of blood cells after oxygen-reduced storage TIFF0007863358000001.tif132170

[0148] Example 7: Platelet deoxygenation does not impair hemostatic performance. Eight units of whole blood (WB) are obtained from healthy, consenting volunteers through standard 500 mL blood donations. The donated whole blood is collected in CPDA-1 anticoagulant (Research Blood Components, Inc.) as described in Example 2, and leukocytes are reduced using a platelet preservation filter (Imuflex® WB-SP) (LRWB; Terumo Medical Corporation). The resulting filtered units are then divided into two. One half of the units is cryogenically stored aerobically as usual, and the second half is further divided and cryogenically stored anaerobically. The anaerobically stored units have reduced oxygen (OR-LRWB) or reduced oxygen and carbon dioxide (OCR-LRWB). The anaerobically stored units are processed using a Sorin D100 membrane oxygenator to obtain anaerobic units containing approximately 5% SO2 and approximately 35 mmHgpCO2. The resulting anaerobic units are placed in standard PVC bags and stored in anaerobic canisters containing oxygen absorbers and nitrogen gas. A pair of leukocyte-reduced platelet units are evaluated weekly throughout the entire 21-day storage period as described below.

[0149] Following the manufacturer's instructions, units were evaluated for metabolic parameters including percent hemolysis (Plasma Low, Angelholm, Sweden), ATP (DiaSys, Flacht, Germany), and 2,3-DPG (Sigma-Aldrich, St. Louis, MO). As shown in Figure 4A, lower levels of ATP were maintained in stored OCR-LRWB compared to conventionally stored LRWB (solid line), but increased in stored OR-LRWB. As shown in Figure 4B, elevated levels of 2,3-DPG were maintained in stored OCR-LRWB and OR-LRWB for up to 21 days compared to conventionally stored LRWB. Furthermore, as shown in Figure 4C, no significant changes in hemolysis were observed when comparing stored OR-LRWB and stored OCR-LRWB to conventionally stored LRWB (solid line).

[0150] Plasma coagulation parameters were evaluated by assessing prothrombin time (PT), activated partial prothrombin time (aPTT), and fibrinogen and D-dimer levels in conventionally preserved LRWB and OCR-LRWB. As shown in Figure 5, aPTT and PT were slightly prolonged in conventionally preserved LRWB (solid line), but not significantly. No evidence of coagulation activation was observed, as is evident from similar fibrinogen and D-dimer levels.

[0151] Plasma coagulation factors were further evaluated by determining the activity levels of factor V, factor VIII, protein C activity, protein S activity, and von Willebrand factor (vWF) in conventionally stored LRWB and OCR-LRWB. Analysis of protein C and protein S was performed using ACL TOP® (Instrumentation Laboratory) and STA-R Evolution Coagulation Analyzer® (Diagnostica Stago, Inc.), respectively, according to the manufacturer's instructions. As shown in Figure 6, no significant changes were observed in the levels of factor V, factor VIII, protein C activity, protein S activity, and vWF in anaerobically cryogenically stored OCR-LRWB (dashed line) compared to conventionally stored WB (solid line).

[0152] Conventional LRWB and OCR-LRWB samples were further evaluated for coagulation using thromboelastography (TEG) equipped with a Haemoscope Thromboelastograph® analyzer (Haemonetics) according to the manufacturer's instructions. As shown in Figures 7A–7D, no significant differences were observed in propagation (TEG angle), amplification (TEG K), maximum amplitude (TEG MA), or reaction time (TEG R) in the OCR-LRWB (dashed line) compared to conventionally stored LRWB (solid line).

[0153] While this disclosure has been described with reference to specific embodiments, those skilled in the art will understand that various modifications may be made and equivalents may be substituted for their elements without departing from the scope of this disclosure. Furthermore, many modifications may be made to adapt the teachings of this disclosure to specific circumstances or materials without departing from the scope of this disclosure.

[0154] Therefore, this disclosure is not limited to any particular embodiment disclosed as the best form intended to carry out this disclosure, but is intended to include all embodiments within the scope and spirit of the appended claims.

Claims

1. Whole blood for transfusion to trauma patients requiring transfusion, having an oxygen saturation (SO2) of 10% or less before and during storage, and containing platelets in an anticoagulant solution, comprising oxygen and carbon dioxide reduced-leukocyte reduced whole blood (OCR-LRWB+PLT), The aforementioned OCR-LRWB+PLT was stored for a maximum of three weeks. The aforementioned stored OCR-LRWB+PLT has one or more coagulation parameters equivalent to or better than the same one or more coagulation parameters found in whole blood (non-OCR-LRWB+PLT) that has been stored in the conventional manner, containing platelets but with reduced oxygen and carbon dioxide levels, and with reduced white blood cell count. The stored whole blood wherein one or more coagulation parameters are selected from the group consisting of thromboelastography angle (TEG angle), thromboelastography blood kinetics (TEG K), thromboelastography maximum amplitude (TEG MA), thromboelastography reaction time (TEG R), prothrombin time (PT), partial thromboplastin time (PTT), fibrinogen level, D-dimer value, and thrombin generation value.

2. The preserved whole blood according to Claim 1, wherein the coagulation parameter is the TEG angle, and the TEG angle exceeds 40°.

3. The preserved whole blood according to claim 1, wherein the coagulation parameter is TEG K, and the TEG K is between 1 minute and 5 minutes.

4. The preserved whole blood according to claim 1, wherein the coagulation parameter is TEG MA, and the TEG MA is between 35 millimeters (mm) and 65 millimeters (mm).

5. The preserved whole blood according to claim 1, wherein the coagulation parameter is TEG R, and the TEG R is between 4 minutes and 8 minutes.

6. The preserved whole blood according to claim 1, wherein the coagulation parameter is PT, and the PT is between 10 seconds and 15 seconds.

7. The stored whole blood according to claim 1, wherein the coagulation parameter is PPT, and the PPT is between 32 seconds and 42 seconds.

8. The stored whole blood according to claim 1, wherein the coagulation parameter is the fibrinogen level, and the fibrinogen level is between 250 milligrams / deciliter (mg / dL) and 350 mg / dL.

9. The stored OCR-LRWB+PLT has a coagulation factor amount greater than or equal to the same amount of coagulation factor in conventionally stored non-OCR-LRWB+PLT. The preserved whole blood according to claim 1, wherein the coagulation factor is selected from the group consisting of factor V, factor VIII, antithrombin (AT), protein C, von Willebrand factor (vWF), and any combination thereof.

10. The preserved whole blood according to claim 1, wherein the preserved OCR-LRWB+PLT exhibits higher red blood cell (RBC) deformability compared to the red blood cell (RBC) deformability of non-OCR-LRWB+PLT preserved in the conventional manner.

11. The stored whole blood according to claim 1, wherein the anticoagulant solution is selected from the group consisting of citrate-phosphate-dextrose (CPD), citrate-phosphate-dextrose-adenine solution 1 (CPDA-1), acidic-citric acid-dextrose (ACD), and acidic-citric acid-dextrose solution A (ACD-A).

12. The stored whole blood according to claim 1, wherein the trauma patient requiring blood transfusion is a patient requiring frequent blood transfusions.

13. The stored whole blood according to claim 1, wherein the trauma patient requiring blood transfusion is a patient with hemorrhagic trauma or a patient with blunt trauma.

14. The preserved whole blood according to claim 1, wherein the preserved OCR-LRWB+PLT is safe to transfuse to trauma patients requiring transfusion after a storage period of at least three weeks.

15. The preserved whole blood according to claim 1, wherein the trauma patient is a transplant patient, a cardiac surgery patient, an obstetric patient, a digestive surgery patient, a cancer patient, or an orthopedic surgery patient.

16. A massive blood transfusion kit containing plasma and oxygen- and carbon dioxide-reduced concentrated red blood cells (OCR-pRBC+PLT) containing platelets, sealed in an impermeable enclosure to maintain a reduced oxygen and carbon dioxide state.

17. The massive blood transfusion kit according to claim 16, wherein the volume ratio of plasma to OCR-pRBC+PLT is 1:

1.

18. The massive blood transfusion kit according to claim 16, wherein the volume ratio of plasma to OCR-pRBC+PLT is 1:

2.

19. The massive blood transfusion kit according to claim 16, further containing platelets, wherein the volume ratio of plasma, platelets, and OCR-pRBC+PLT is 1:1:

1.

20. The massive blood transfusion kit according to claim 16, further containing platelets, wherein the volume ratio of plasma, platelets, and OCR-pRBC+PLT is 1:1:

2.

21. The whole blood according to claim 1, wherein the OCR-LRWB+PLT has an oxygen saturation (SO₂) of 5% or less before and after storage, and a partial pressure of carbon dioxide of 35 millimeters of mercury (mmHg) or less.

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