WO2020047157A1

WO2020047157A1 - Citrate-free platelet additive solutions, compositions and methods thereof

Info

Publication number: WO2020047157A1
Application number: PCT/US2019/048653
Authority: WO
Inventors: Todd GETZ
Original assignee: American National Red Cross
Priority date: 2018-08-31
Filing date: 2019-08-28
Publication date: 2020-03-05

Abstract

Described herein are citrate-free platelet additive solutions, platelet compositions containing citrate-free platelet additive solutions, and methods of making and using the same.

Description

CITRATE-FREE PLATELET ADDITIVE SOLUTIONS, COMPOSITIONS AND

METHODS THEREOF

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No.

62/725,406, filed on August 31, 2018, which is incorporated by reference herein in its entirety.

FIELD OF THE INVENTION

[0002] The present invention relates to citrate-free platelet additive solutions, platelet compositions containing citrate-free platelet additive solutions, and methods of making and using the same.

BACKGROUND OF THE INVENTION

[0003] Platelets are a component of blood whose function is to react to bleeding by clumping and forming blood clots. Platelets can be isolated from whole blood or by a process called platelet apheresis where blood is taken from a donor, passed through a device which removes the platelets, and the remainder is returned to the donor.

[0004] Isolated platelets can be stored in blood plasma. However, because plasma may cause immune-mediated transfusion reactions, platelet storage solutions (also called platelet additive solutions) were developed to reduce the amount of plasma per transfusion and reduce the frequency and/or severity of transfusion reactions.

[0005] Citrate is commonly used in platelet additive solutions because it chelates calcium and prevents spontaneous aggregation of platelets. Mollison et al ., Br. J. Haematol. 2000; 108(1): 13-18; Van Rhenen et al ., Transfusion. 1995; 35(10): 50-53 ; Ringwald et al ., Transfus. Med. Rev. 2006; 20(2): 158-64. A ratio of three molecules of citrate to one molecule of calcium has been reported to prevent clotting. Mollison et al ., Br. J. Haematol. 2000; 108(1): 13-18. A 5: 1 ratio of whole blood to 3.8% citrate is also used in practice. This ratio may have been adopted from O.H. Robertson who developed a transfusion bottle in the early l900's that utilized 3.8% citrate solution. Mollison et al ., Br. J. Haematol. 2000; 108(1): 13-18. The final concentration of citrate in Robertson's bottle was ~l0mM/L. It has been suggested that citrate should remain in platelet additive solutions because a citrate concentration below 8mM/L may trigger spontaneous clotting. Gulliksson, Transfusion 1993; 33:301-303.

[0006] Concentrations of citrate between 5-l0mM/L have also been reported to induce apoptosis in a variety of oncogenic cell lines. Lu et al., Anticancer Res. 2011; 31(3):797- 805; Kruspig et al., Cell. Mol. Life Sci. 2012; 69(24):4229-37. While citrate has been shown to suppress glycolysis through inhibition of phosphofructokinase, several investigators have reported a different mechanism of cell death which is mediated through a mitochondrial pathway involving caspase activation. Lu et al., Anticancer Res. 2011; 3 l(3):797-805; Kruspig et al, Cell. Mol. Life Sci. 2012; 69(24):4229-37.

[0007] Despite the development of platelet additive solutions, platelets have a relatively short shelf life due to a progressive decline in platelet viability and function over time. As such, regulations from the U.S. Food and Drug Administration (FDA) indicate that platelets stored between 20-24°C should be used within 5 days from the day of collection, with some limited exceptions for use within 7 days.

[0008] Pathogen reduction (or pathogen reduction system) is a process of treating platelets or other blood products with a photoactive compound that specifically targets DNA or RNA. Examples include, but are not limited to, ultraviolet light exposure and/or reagents that inhibit DNA or RNA replication (e.g., ultraviolet A light and/or a psoralen). The only pathogen reduction system currently approved by the FDA is the Intercept® blood system which utilizes platelets stored in a platelet additive solution containing lOmM sodium citrate. However, data suggests that pathogen reduction processes impair platelet quality and function. Van Aelst et al. Vox Sang. 2015; l08(4):328-39; Stivala et al. Haematologica. 2017; 102(10): 1650-60.

[0009] Improved platelet additive solutions are needed. SUMMARY OF THE INVENTION

[0010] In some embodiments, the present invention relates to a platelet additive solution comprising phosphate, acetate and chloride, wherein the solution does not contain citrate. In some embodiments, the phosphate is sodium phosphate. In some embodiments, the sodium phosphate is monobasic sodium phosphate. In some embodiments, the sodium phosphate is dibasic sodium phosphate. In some embodiments, the dibasic sodium phosphate is anhydrous dibasic sodium phosphate. In some embodiments, the sodium phosphate is a combination of monobasic sodium phosphate and dibasic sodium phosphate. In some embodiments, the acetate is sodium acetate. In some embodiments, the chloride is sodium chloride.

[0011] In other embodiments, the present invention relates to a platelet additive solution comprising sodium phosphate, sodium acetate, and sodium chloride, wherein the solution does not contain citrate. In some embodiments, the platelet additive solution comprises from about 5mM to about 40mM sodium phosphate, from about 20mM to about 50mM sodium acetate, and from about 50mM to about 90mM sodium chloride. In some embodiments, the platelet additive solution comprises from about 20mM to about 30mM sodium phosphate, from about 25mM to about 40mM sodium acetate, and from about 70mM to about 80mM sodium chloride. In some embodiments, the platelet additive solution comprises from about 25mM to about 30mM sodium phosphate, from about 25mM to about 35mM sodium acetate, and from about 70mM to about 80mM sodium chloride.

[0012] In other embodiments, the present invention relates to a platelet additive solution comprising monobasic sodium phosphate, dibasic sodium phosphate, sodium acetate, and sodium chloride, wherein the solution does not contain citrate. In some embodiments, the dibasic sodium phosphate is anhydrous dibasic sodium phosphate. In some embodiments, the platelet additive solution comprises from about lmM to about l5mM monobasic sodium phosphate, from about lOmM to about 30mM dibasic sodium phosphate, from about 20mM to about 50mM sodium acetate, and from about 50mM to about 90mM sodium chloride. In some embodiments, the platelet additive solution comprises from about 5mM to about lOmM monobasic sodium phosphate, from about l5mM to about 25mM dibasic sodium phosphate, from about 20mM to about 40mM sodium acetate, and from about 60mM to about 80mM sodium chloride. In some embodiments, the platelet additive solution comprises from about 5mM to about lOmM monobasic sodium phosphate, from about 20mM to about 25mM dibasic sodium phosphate, from about 30mM to about 35mM sodium acetate, and from about 70mM to about 80mM sodium chloride.

[0013] In other embodiments, the present invention relates to a platelet additive solution comprising potassium, magnesium, glucose, calcium, hispidin, zinc, or a combination thereof. In some embodiments, the platelet additive solution comprises from about lmM to about lOmM potassium, from about O.lmM to about 5mM magnesium, from about lmM to about 20mM glucose, from about O.OlmM to about 4mM calcium, or a combination thereof.

[0014] In other embodiments, the present invention relates to a platelet additive solution comprising sodium phosphate, sodium acetate, sodium chloride, potassium chloride, magnesium chloride and glucose, wherein the solution does not contain citrate. In some embodiments, the platelet additive solution comprises from about lOmM to about 40mM sodium phosphate, from about 20mM to about 50mM sodium acetate, from about 50mM to about 90mM sodium chloride, from about lmM to about lOmM potassium chloride, from about O.lmM to about 5mM magnesium chloride, and from about lmM to about 20mM glucose. In some embodiments, the platelet additive solution comprises from about 20mM to about 30mM sodium phosphate, from about 20mM to about 40mM sodium acetate, from about 60mM to about 70mM sodium chloride, from about lmM to about 5mM potassium chloride, from about lmM to about 2mM magnesium chloride, and from about lOmM to about 20mM glucose. In some embodiments, the platelet additive solution comprises from about 5mM to about lOmM monobasic sodium phosphate, from about l5mM to about 25mM dibasic sodium phosphate, from about lOmM to about 40mM sodium acetate, from about 50mM to about 90mM sodium chloride, from about lmM to about lOmM potassium chloride, from about O. lmM to about 5mM magnesium chloride, and from about lmM to about 20mM glucose. In some embodiments, the dibasic sodium phosphate is anhydrous dibasic sodium phosphate. In some embodiments, the platelet additive solution comprises from about 5mM to about lOmM monobasic sodium phosphate, from about 20mM to about 25mM dibasic sodium phosphate, from about 20mM to about 40mM sodium acetate, from about 60mM to about 80mM sodium chloride, from about lmM to about 5mM potassium chloride, from about lmM to about 5mM magnesium chloride, and from about lOmM to about 20mM glucose.

[0015] In some embodiments, the platelet additive solution has a pH of about 6.2 or greater.

[0016] In some embodiments, the platelet additive solution preserves platelet function, activation, or a combination thereof.

[0017] In some embodiments, the platelet additive solution does not cause spontaneous clotting of platelets.

[0018] In some embodiments, the platelet additive solution results in one or more of the following when used to store platelets:

(i) decreased levels of reactive oxygen species,

(ii) decreased glucose utilization,

(iii) decreased lactate production,

(iv) decreased P-selectin expression,

(v) decreased apoptosis,

(vi) increased aggregation to collagen,

(vii) increased aggregation to TRAP6,

(viii) increased cell function, and

(ix) increased cell triggered activation,

compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0019] In some embodiments, the platelet additive solution results in one or more of the following when used to store platelets:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation,

compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate. [0020] In some embodiments, the platelet additive solution results in one or more of the following when used to store platelets:

(i) a decrease of from about 30% to about 60% in reactive oxygen species levels,

(ii) a decrease of from about 30% to about 60% in cell death,

(iii) a decrease of from about 40% to about 80% in glucose utilization,

(iv) a decrease of from about 30% to about 60% in lactate production,

(v) a decrease of from about 20% to about 50% in CD62P expression,

(vi) an increase of from about 10% to about 50% in collagen aggregation, and

(vii) an increase of from about 10% to about 50% in TRAP6 aggregation,

[0021] In some embodiments, the citrate in the reference platelet additive solution is from about lmM citrate to about 20mM citrate.

[0022] In some embodiments, the platelet additive solution is for storage of platelets for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, or at least 10 days. In some embodiments, the platelet additive solution is for storage of platelets from about 1 day to about 10 days, from about 1 day to about 7 days, or from about 1 day to about 5 days. In some embodiments, the platelet additive solution is for storage of platelets for about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or about 10 days. In some embodiments, the platelets are platelets treated with a pathogen reduction system. In some embodiments, the pathogen reduction system is ultraviolet light exposure and/or a reagent that inhibits the replication of DNA or RNA.

[0023] In other embodiments, the present invention relates to a composition comprising a platelet additive solution described herein and platelets. In some embodiments, the platelets are platelets treated with a pathogen reduction system. In some embodiments, the pathogen reduction system is ultraviolet light exposure and/or a reagent that inhibits the replication of DNA or RNA.

[0024] In other embodiments, the present invention relates to methods of using and making a platelet additive solution described herein or a platelet composition described herein. For example, in some embodiments, the present invention relates to a method for storing platelets comprising (i) combining platelets with a platelet additive solution described herein or providing a platelet composition described herein, and (ii) storing the combination of (i) or composition for at least 1 day, at least 2 days, at least 3 days, at least

4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, or at least 10 days. In some embodiments, the platelets exhibit one or more of the following after storage:

(i) decreased levels of reactive oxygen species,

(ii) decreased glucose utilization,

(iii) decreased lactate production,

(iv) decreased P-selectin expression,

(v) decreased apoptosis,

(vi) increased aggregation to collagen,

(vii) increased aggregation to TRAP6,

(viii) increased cell function, and

(ix) increased cell triggered activation,

[0025] In some embodiments, the platelets exhibit one or more of the following after storage:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation,

[0026] In some embodiments, the platelets exhibit one or more of the following after storage:

(ii) a decrease of from about 30% to about 60% in cell death, (iii) a decrease of from about 40% to about 80% in glucose utilization,

(iv) a decrease of from about 30% to about 60% in lactate production,

(v) a decrease of from about 20% to about 50% in CD62P expression,

(vi) an increase of from about 10% to about 50% in collagen aggregation, and

(vii) an increase of from about 10% to about 50% in TRAP6 aggregation,

[0027] In other embodiments, the present invention is directed to a method for decreasing platelet activation or aggregation during storage, comprising (i) combining platelets with a platelet additive solution of the invention or providing a platelet composition of the invention, and (ii) storing the combination or composition of (i) for least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, or at least 10 days. In some embodiments, the platelets have (i) a decrease of at least about 30% in reactive oxygen species levels, (ii) a decrease of at least about 40% in glucose utilization, (iii) a decrease of at least about 30% in lactate production, (iv) a decrease of at least about 20% in CD62P expression, or (v) a combination thereof, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate. In some embodiments, the platelets have (i) a decrease of from about 30% to about 60% in reactive oxygen species levels, (ii) a decrease of from about 40% to about 80% in glucose utilization, (iii) a decrease of from about 30% to about 60% in lactate production, (iv) a decrease of from about 20% to about 50% in CD62P expression, or (v) a combination thereof, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0028] In other embodiments, the present invention is directed to a method for increasing triggered platelet activation or aggregation, comprising (i) combining platelets with a platelet additive solution of the invention or providing a platelet composition of the invention, and (ii) storing the combination or composition of (i) for least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, or at least 10 days. In some embodiments, the platelets have an increase of at least about 10% activation or aggregation, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate. In some embodiments, the platelets have (i) a decrease of at least about 20% in CD62P expression, (ii) an increase of at least about 10% in collagen aggregation, (iii) an increase of at least about 10% in TRAP6 aggregation, or (iv) a combination thereof, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate. In some embodiments, the platelets have (i) a decrease of from about 20% to about 50% in CD62P expression, (ii) an increase of from about 10% to about 50% in collagen aggregation, (iii) an increase of from about 10% to about 50% in TRAP6 aggregation, or (iv) a combination thereof, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0029] In other embodiments, the present invention is directed to a method of making a platelet additive solution, comprising combining phosphate, acetate and chloride to form a solution, wherein the solution does not contain citrate. In some embodiments, the phosphate is sodium phosphate, the acetate is sodium acetate, and the chloride is sodium chloride. In some embodiments, the sodium phosphate is a combination of monobasic sodium phosphate and dibasic sodium phosphate. In some embodiments, the dibasic sodium phosphate is anhydrous dibasic sodium phosphate.

[0030] In other embodiments, the present invention is directed to a method of making a platelet additive solution, comprising combining sodium phosphate, sodium acetate, sodium chloride, potassium chloride, magnesium chloride, and glucose to form a solution, wherein the solution does not contain citrate. In some embodiments, the sodium phosphate is a combination of monobasic sodium phosphate and dibasic sodium phosphate.

[0031] In other embodiments, the present invention is directed to a method of treating or preventing hemorrhage in a subject in need thereof, comprising administering a platelet composition of the invention to the subject. In some embodiments, the hemorrhage is acute or severe hemorrhage.

[0032] In other embodiments, the present invention is directed to a method of treating or preventing low platelet count in a subject in need thereof, comprising administering a platelet composition of the invention to the subject. In some embodiments, the subject is undergoing chemotherapy.

[0033] In some embodiments of the methods of the invention, the subject is a human.

[0034] In some embodiments of the methods of the invention, the platelets are stored from about 1 day to about 10 days, from about 1 day to about 7 days, or from about 1 day to about 5 days. In some embodiments, the platelets are stored for about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or about 10 days.

[0035] In some embodiments of the methods of the invention, the citrate in the reference platelet additive solution is from about lmM citrate to about 20mM citrate.

[0036] In some embodiments of the methods of the invention, the platelets are treated with a pathogen reduction system. In some embodiments, the pathogen reduction system is ultraviolet light exposure and/or a reagent that inhibits the replication of DNA or RNA.

[0037] In other embodiments, the present invention is directed to a method for reducing pathogens in a platelet composition, comprising (i) combining platelets with a platelet additive solution of the invention or providing a platelet composition of the invention, and

(ii) storing the combination or composition of (i) for least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 6 days, at least 9 days, or at least 10 days. In some embodiments, the platelet composition has at least 10% fewer pathogens after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0038] In other embodiments, the present invention is directed to a method for decreasing platelet activation or aggregation during storage or increasing triggered platelet activation or aggregation, comprising (i) combining platelets with a platelet additive solution of the invention or providing a platelet composition of the invention, and (ii) storing the combination or composition of (i) for least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 6 days, at least 9 days, or at least 10 days. In some embodiments, the platelets exhibit one of more of the following after storage:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Some embodiments of the invention are described herein, by way of example only, with reference to the accompanying drawings. With specific reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of embodiments of the invention.

[0040] FIG. 1 A shows reactive oxygen levels (mean ± standard deviation (SD)) following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or lOmM citrate using the CM-H2DCFDA probe. * = P = <0.05 statistical significance from four independent experiments by one-way ANOVA.

[0041] FIG. 1B shows annexin V levels (mean ± SD) following storage of platelets for 1,

5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or lOmM citrate. * = P = <0.05 statistical significance from four independent experiments by one- way ANOVA.

[0042] FIG. 2A shows glucose levels (mean ± SD) following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or lOmM citrate.

* = P = <0.05 statistical significance from four independent experiments by one-way ANOVA. NS = not statistically significant.

[0043] FIG. 2B shows lactate levels (mean ± SD) following storage of platelets for 1, 5 or

7 days in test platelet additive solutions containing OmM, 2mM, 5mM or 10 mM citrate.

* = P = <0.05 statistical significance from four independent experiments by one-way ANOVA.

[0044] FIG. 2C shows pH levels (mean ± SD) following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or 10 mM citrate. * = P = <0.05 statistical significance from four independent experiments by one-way ANOVA.

[0045] FIG. 3A shows levels (mean ± SD) of the platelet surface marker CD62P (P- selectin) following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or lOmM citrate. * = P = <0.05 statistical significance from four independent experiments by one-way ANOVA.

[0046] FIG. 3B shows levels (mean ± SD) of the platelet surface receptor CD42b (GPIb) following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or lOmM citrate. Data is from four independent experiments. NS = not statistically significant.

[0047] FIG. 4A shows aggregation data (mean ± standard error of the mean (SEM)) of platelets stored in test platelet additive solutions containing OmM or lOmM citrate for 1, 5 or 7 days (Dl, D5 and D7, respectively). Aggregation was evaluated in response to stimulation with collagen in Tyrode's buffer ("Tyrodes"). Data is from three independent experiments.

[0048] FIG. 4B shows aggregation data (mean ± SEM) of platelets stored in test platelet additive solutions containing OmM or lOmM citrate for 1, 5 or 7 days (Dl, D5 and D7, respectively). Aggregation was evaluated in response to stimulation with TRAP6 ("TRAP") in Tyrode's buffer ("Tyrodes"). Data is from three independent experiments.

[0049] FIG. 4C shows aggregation data (mean ± SEM) of platelets stored in test platelet additive solutions containing OmM or lOmM citrate for 1, 5 or 7 days (Dl, D5 and D7, respectively). Aggregation was evaluated in response to stimulation with collagen in plasma. Data is from three independent experiments.

[0050] FIG. 4D shows aggregation data (mean ± SEM) of platelets stored in test platelet additive solutions containing OmM or lOmM citrate for 1, 5 or 7 days (Dl, D5 and D7, respectively). Aggregation was evaluated in response to stimulation with TRAP6 ("TRAP") in plasma. Data is from three independent experiments.

[0051] FIG. 5A shows reactive oxygen levels (mean ± SEM) following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate) using the CM-H2DCFDA probe. * = P = <0.05 statistical significance from seven independent experiments by paired T-test.

[0052] FIG. 5B shows annexin V levels (mean ± SEM) following storage of platelets for

1, 5 or 7 days in citrate-free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate). * = P = <0.05 statistical significance from seven independent experiments by paired T- test.

[0053] FIG. 6A shows glucose levels (mean ± SEM) following storage of platelets for 1,

5 or 7 days in citrate-free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate). * = P = <0.05 statistical significance from seven independent experiments by paired T-test. NS = no statistical significance. [0054] FIG. 6B shows lactate levels (mean ± SEM) following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate). * = P = <0.05 statistical significance from seven independent experiments by paired T-test. NS = no statistical significance.

[0055] FIG. 6C shows pH levels (mean ± SEM) following storage of platelets for 1, 5 or

7 days in citrate-free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate). * = P = <0.05 statistical significance from seven independent experiments by paired T-test.

[0056] FIG. 7A shows levels (mean ± SEM) of CD62P (P-selectin) following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate). * = P = <0.05 statistical significance from seven independent experiments by paired T-test.

[0057] FIG. 7B shows levels (mean ± SEM) of CD42b (GPIb) following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate). NS = no statistical significance.

[0058] FIG. 8A shows platelet aggregation (mean ± SEM) following storage in citrate- free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate) in response to stimulation with collagen in plasma. * = P = <0.05 statistical significance from seven independent experiments by paired T-test. NS = no statistical significance.

[0059] FIG. 8B shows platelet aggregation (mean ± SEM) following storage in citrate- free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate) in response to stimulation with collagen in Tyrode's buffer ("Tyrodes"). * = P = <0.05 statistical significance from seven independent experiments by paired T-test.

[0060] FIG. 8C shows platelet aggregation (mean ± SEM) following storage in citrate- free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate) in response to stimulation with TRAP6 in plasma. * = P = <0.05 statistical significance from seven independent experiments by paired T-test.

[0061] FIG. 8D shows platelet aggregation (mean ± SEM) following storage in citrate- free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate) in response to stimulation with TRAP6 in Tyrode's buffer ("Tyrodes"). * = P = <0.05 statistical significance from seven independent experiments by paired T-test..

[0062] FIG. 8E shows platelet aggregation (mean ± SEM) following storage in citrate- free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate) in response to stimulation with collagen and adenosine diphosphate (ADP) in plasma. * = P = <0.05 statistical significance from seven independent experiments by paired T-test.

[0063] FIG. 8F shows platelet aggregation (mean ± SEM) following storage of in citrate- free PAS-3 (0 mM Citrate) or InterSol solution (10 mM Citrate) in response to stimulation with collagen and ADP in Tyrode's buffer ("Tyrodes"). * = P = <0.05 statistical significance from seven independent experiments by paired T-test.

[0064] FIGs. 9 A and 9C show aggregation data of platelets stored in PAS 5 A or PAS-P1 for 1 day. Aggregation was evaluated in response to stimulation with collagen. Data is from a single experiment.

[0065] FIGs. 9B and 9D show aggregation data of platelets stored in PAS 5 A or PAS-P1 for 1 day. Aggregation was evaluated in response to stimulation with TRAP6. Data is from a single experiment.

[0066] FIGs. 10A and 10C show aggregation data of platelets stored in PAS 5 A or PAS-

Pl for 5 days. Aggregation was evaluated in response to stimulation with collagen or TRAP6. Data is from a single experiment.

[0067] FIGs. 10B and 10D show aggregation data of platelets stored in PAS 5 A or PAS-

[0068] FIGs. 11A and 11C show aggregation data of platelets stored in PAS 5A or PAS-

Pl for 8 days. Aggregation was evaluated in response to stimulation with collagen or TRAP6. Data is from a single experiment.

[0069] FIGs. 11B and 11D show aggregation data of platelets stored in PAS 5A or PAS-

[0070] FIG. 12 shows platelet count, blood gas and flow cytometry measurements of platelets following pathogen reduction and storage in citrate-free PAS-3 or InterSol solution (10 mM Citrate) for 0, 1, 2 and 4 days. Data is from one experiment.

[0071] FIG. 13 A shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 0, pre-illumination or prior to exposure of ultraviolet A (UVA) treatment. Aggregation was evaluated in response to stimulation with collagen in Tyrode's buffer. Data is from one experiment. [0072] FIG. 13B shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 0, pre-illumination or prior to exposure of UVA treatment. Aggregation was evaluated in response to stimulation with TRAP6 in Tyrode's buffer. Data is from one experiment.

[0073] FIG. 13C shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 0, pre-illumination or prior to exposure of UVA treatment. Aggregation was evaluated in response to stimulation with collagen and adenosine diphosphate (ADP) in Tyrode's buffer. Data is from one experiment.

[0074] FIG. 14A shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 0, post-illumination or exposure to UVA treatment. Aggregation was evaluated in response to stimulation with collagen in Tyrode's buffer. Data is from one experiment.

[0075] FIG. 14B shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 0, post-illumination or exposure to UVA treatment. Aggregation was evaluated in response to stimulation with TRAP6 in Tyrode's buffer. Data is one from experiment.

[0076] FIG. 14C shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 0, post-illumination or exposure to UVA treatment. Aggregation was evaluated in response to stimulation with collagen and ADP in Tyrode's buffer. Data is from one experiment.

[0077] FIG. 15A shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 1, post-illumination or exposure to UVA treatment. Aggregation was evaluated in response to stimulation with collagen in Tyrode's buffer. Data is from one experiment.

[0078] FIG. 15B shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 1, post-illumination or exposure to UVA treatment. Aggregation was evaluated in response to stimulation with TRAP6 in Tyrode's buffer. Data is from one experiment.

[0079] FIG. 15C shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 1, post-illumination or exposure to UVA treatment. Aggregation was evaluated in response to stimulation with collagen and ADP in Tyrode's buffer. Data is from one experiment. [0080] FIG. 16A shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 4, post-illumination or exposure to UVA treatment. Aggregation was evaluated in response to stimulation with collagen in Tyrode's buffer. Data is from one experiment.

[0081] FIG. 16B shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 4, post-illumination or exposure to UVA treatment. Aggregation was evaluated in response to stimulation with TRAP6 in Tyrode's buffer. Data is from one experiment.

[0082] FIG. 16C shows aggregation data of platelets stored in citrate-free PAS-3 or

InterSol solution at day 4, post-illumination or exposure to UVA treatment. Aggregation was evaluated in response to stimulation with collagen and ADP in Tyrode's buffer. Data is from one experiment.

DETAILED DESCRIPTION OF THE INVENTION

[0083] Before explaining embodiments of the invention in detail, it is to be understood that the invention is not limited in its application to the details set forth in the following description or exemplified by the Examples. The invention is capable of other embodiments or of being practiced or carried out in various ways. Also, it is to be understood that the phraseology and terminology employed herein is for the purpose of description and should not be regarded as limiting.

General Definitions

[0084] As used herein, the terms "comprises", "comprising", "includes", "including",

"having", and their conjugates mean "including but not limited to."

[0085] As used herein, the term "consisting of' means "including and limited to."

[0086] As used herein, the term "consisting essentially of' means the specified material of a composition, or the specified steps of a method, and those additional materials or steps that do not materially affect the basic characteristics of the material or method.

[0087] As used herein, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof. [0088] As used herein, the term "about" modifying an amount related to the invention refers to variation in the numerical quantity that can occur, for example, through routine testing and handling; through inadvertent error in such testing and handling; through differences in the manufacture, source, or purity of ingredients employed in the invention; and the like. Whether or not modified by the term "about", the claims include equivalents of the recited quantities. In one embodiment, the term "about" means within 10% of the reported numerical value. In another embodiment, the term "about" means within 5% of the reported numerical value.

[0089] As used herein, the term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

[0090] As used herein, the term "treating" includes abrogating, substantially inhibiting, slowing or reversing the progression of a condition, substantially ameliorating clinical or aesthetical symptoms of a condition or substantially preventing the appearance of clinical or aesthetical symptoms of a condition.

[0091] As used herein, the term "platelet additive solution" refers to a storage media for platelets.

[0092] As used herein, the term "citrate" refers to any derivative ( e.g ., salts, acid salts, esters, or polyatomic anions) of citric acid. Examples of citrate include, but are not limited to, sodium citrate (e.g., monosodium citrate, disodium citrate, trisodium citrate), triethyl citrate, or a combination thereof.

[0093] As used herein, the terms "citrate-free", "does not contain citrate" or "does not comprise citrate" are used interchangeably and refer to a platelet additive solution or platelet composition described herein that contains no citrate.

[0094] As used herein, the term "phosphate" refers to any derivative of phosphoric acid.

Examples of phosphate include, but are not limited to, sodium phosphate, monobasic sodium phosphate, dibasic sodium phosphate, or a combination thereof.

[0095] As used herein, the term "acetate" refers to any salt formed by the combination of acetic acid with an alkaline, earthy, metallic or nonmetallic base. Examples of acetate include, but are not limited to, hydrogen acetate, sodium acetate, ethyl acetate, potassium acetate, aluminum acetate, ammonium acetate, or a combination thereof.

[0096] As used herein, the term "chloride" refers to any chemical compound in which one or more chloride (Cl) ions are covalently bonded. Examples of chloride include, but are not limited to, sodium chloride, potassium chloride, magnesium chloride, calcium chloride, or a combination thereof.

[0097] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.

[0098] Throughout this application, various embodiments of this invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range, such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5 and 6. This applies regardless of the breadth of the range.

Platelet Additive Solutions and Platelet Compositions

[0099] In some embodiments, the present invention relates to citrate-free platelet additive solutions. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of phosphate, acetate and chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of phosphate, acetate and chloride, wherein the solution does not contain citrate. [0100] In some embodiments, the phosphate of the platelet additive solution is sodium phosphate. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of sodium phosphate, acetate and chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of sodium phosphate, acetate and chloride, wherein the solution does not contain citrate.

[0101] In some embodiments, the sodium phosphate of the platelet additive solution is monobasic sodium phosphate. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of monobasic sodium phosphate, acetate and chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of monobasic sodium phosphate, acetate and chloride, wherein the solution does not contain citrate.

[0102] In some embodiments, the sodium phosphate of the platelet additive solution is dibasic sodium phosphate. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of dibasic sodium phosphate, acetate and chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of dibasic sodium phosphate, acetate and chloride, wherein the solution does not contain citrate.

[0103] In some embodiments, the sodium phosphate of the platelet additive solution is a combination of monobasic sodium phosphate and dibasic sodium phosphate. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of a combination of monobasic sodium phosphate and dibasic sodium phosphate, acetate and chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of a combination of monobasic sodium phosphate and dibasic sodium phosphate, acetate and chloride, wherein the solution does not contain citrate.

[0104] In some embodiments, the acetate of the platelet additive solution is sodium acetate. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of phosphate, sodium acetate and chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of phosphate, sodium acetate and chloride, wherein the solution does not contain citrate.

[0105] In some embodiments, the chloride of the platelet additive solution is sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of phosphate, acetate and sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of phosphate, acetate and sodium chloride, wherein the solution does not contain citrate.

[0106] In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of sodium phosphate, sodium acetate and chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of sodium phosphate, sodium acetate and chloride, wherein the solution does not contain citrate. In some embodiments, the sodium phosphate is monobasic sodium phosphate, dibasic sodium phosphate, or a combination of monobasic sodium phosphate and dibasic sodium phosphate.

[0107] In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of sodium phosphate, acetate and sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of sodium phosphate, acetate and sodium chloride, wherein the solution does not contain citrate. In some embodiments, the sodium phosphate is monobasic sodium phosphate, dibasic sodium phosphate, or a combination of monobasic sodium phosphate and dibasic sodium phosphate.

[0108] In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of phosphate, sodium acetate and sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of phosphate, sodium acetate and sodium chloride, wherein the solution does not contain citrate. In some embodiments, the phosphate is sodium phosphate. In some embodiments, the sodium phosphate is monobasic sodium phosphate, dibasic sodium phosphate, or a combination of monobasic sodium phosphate and dibasic sodium phosphate.

[0109] In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of sodium phosphate, sodium acetate and sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of sodium phosphate, sodium acetate and sodium chloride, wherein the solution does not contain citrate. In some embodiments, the sodium phosphate is monobasic sodium phosphate, dibasic sodium phosphate, or a combination of monobasic sodium phosphate and dibasic sodium phosphate. [0110] In some embodiments, the amount of phosphate in the platelet additive solution is at least about 5mM, at least about lOmM, at least about l5mM, at least about 20mM, at least about 25mM, at least about 30mM, at least about 35mM, at least about 40mM, at least about 45mM, or at least about 50mM. In some embodiments, the amount of phosphate in the platelet additive solution is from about 5mM to about 50mM or any range of values thereof ( e.g ., from about 5mM to about 45mM, from about 5mM to about 40mM, from about 5mM to about 35mM, from about 5mM to about 30mM, from about 5mM to about 25mM, from about 5mM to about 20mM, from about 5mM to about l5mM, from about 5mM to about lOmM, from about lOmM to about 50mM, from about lOmM to about 45mM, from about lOmM to about 40mM, from about lOmM to about 35mM, from about lOmM to about 30mM, from about lOmM to about 25mM, from about lOmM to about 20mM, from about lOmM to about l5mM, from about l5mM to about 50mM, from about l5mM to about 45mM, from about l5mM to about 40mM, from about l5mM to about 35mM, from about l5mM to about 30mM, from about l5mM to about 25mM, from about l5mM to about 20mM, from about 20mM to about 50mM, from about 20mM to about 45mM, from about 20mM to about 40mM, from about 20mM to about 35mM, from about 20mM to about 30mM, from about 20mM to about 25mM, from about 25mM to about 50mM, from about 25mM to about 45mM, from about 25mM to about 40mM, from about 25mM to about 35mM, from about 25mM to about 30mM, from about 30mM to about 50mM, from about 30mM to about 45mM, from about 30mM to about 40mM, from about 30mM to about 35mM, from about 35mM to about 50mM, from about 35mM to about 45mM, from about 35mM to about 40mM, from about 40mM to about 50mM, from about 40mM to about 45mM, or from about 45mM to about 50mM). In some embodiments, the amount of phosphate in the platelet additive solution is about 5mM, about lOmM, about l5mM, about 20mM, about 2lmM, about 22mM, about 23mM, about 24mM, about 25mM, about 26mM, about 27mM, about 28mM, about 29mM, about 30mM, about 35mM, about 40mM, about 45mM, or about 50mM. In some embodiments, the amount of phosphate in the platelet additive solution is about 28.2mM or about 28.3mM.

[0111] In some embodiments, the amount of sodium phosphate in the platelet additive solution is at least about 5mM, at least about lOmM, at least about l5mM, at least about 20mM, at least about 25mM, at least about 30mM, at least about 35mM, at least about 40mM, at least about 45mM, or at least about 50mM. In some embodiments, the amount of sodium phosphate in the platelet additive solution is from about 5mM to about 50mM or any range of values thereof ( e.g ., from about 5mM to about 45mM, from about 5mM to about 40mM, from about 5mM to about 35mM, from about 5mM to about 30mM, from about 5mM to about 25mM, from about 5mM to about 20mM, from about 5mM to about l5mM, from about 5mM to about lOmM, from about lOmM to about 50mM, from about lOmM to about 45mM, from about lOmM to about 40mM, from about lOmM to about 35mM, from about lOmM to about 30mM, from about lOmM to about 25mM, from about lOmM to about 20mM, from about lOmM to about l5mM, from about l5mM to about 50mM, from about l5mM to about 45mM, from about l5mM to about 40mM, from about l5mM to about 35mM, from about l5mM to about 30mM, from about l5mM to about 25mM, from about l5mM to about 20mM, from about 20mM to about 50mM, from about 20mM to about 45mM, from about 20mM to about 40mM, from about 20mM to about 35mM, from about 20mM to about 30mM, from about 20mM to about 25mM, from about 25mM to about 50mM, from about 25mM to about 45mM, from about 25mM to about 40mM, from about 25mM to about 35mM, from about 25mM to about 30mM, from about 30mM to about 50mM, from about 30mM to about 45mM, from about 30mM to about 40mM, from about 30mM to about 35mM, from about 35mM to about 50mM, from about 35mM to about 45mM, from about 35mM to about 40mM, from about 40mM to about 50mM, from about 40mM to about 45mM, or from about 45mM to about 50mM). In some embodiments, the amount of sodium phosphate in the platelet additive solution is about 5mM, about lOmM, about l5mM, about 20mM, about 2lmM, about 22mM, about 23mM, about 24mM, about 25mM, about 26mM, about 27mM, about 28mM, about 29mM, about 30mM, about 3 lmM, about 32mM, about 33mM, about 34mM, about 35mM, about 40mM, about 45mM, or about 50mM. In some embodiments, the amount of sodium phosphate in the platelet additive solution is about 28.2mM or about 28.3mM.

[0112] In some embodiments, the amount of dibasic sodium phosphate in the platelet additive solution is at least about 5mM, at least about lOmM, at least about l5mM, at least about 20mM, at least about 25mM, at least about 30mM, at least about 35mM, at least about 40mM, at least about 45mM, or at least about 50mM. In some embodiments, the amount of dibasic sodium phosphate in the platelet additive solution is from about 5mM to about 50mM or any range of values thereof (e.g., from about 5mM to about 45mM, from about 5mM to about 40mM, from about 5mM to about 35mM, from about 5mM to about 30mM, from about 5mM to about 25mM, from about 5mM to about 20mM, from about 5mM to about l5mM, from about 5mM to about lOmM, from about lOmM to about 50mM, from about lOmM to about 45mM, from about lOmM to about 40mM, from about lOmM to about 35mM, from about lOmM to about 30mM, from about lOmM to about 25mM, from about lOmM to about 20mM, from about lOmM to about l5mM, from about l5mM to about 50mM, from about l5mM to about 45mM, from about l5mM to about 40mM, from about l5mM to about 35mM, from about l5mM to about 30mM, from about l5mM to about 25mM, from about l5mM to about 20mM, from about 20mM to about 50mM, from about 20mM to about 45mM, from about 20mM to about 40mM, from about 20mM to about 35mM, from about 20mM to about 30mM, from about 20mM to about 25mM, from about 25mM to about 50mM, from about 25mM to about 45mM, from about 25mM to about 40mM, from about 25mM to about 35mM, from about 25mM to about 30mM, from about 30mM to about 50mM, from about 30mM to about 45mM, from about 30mM to about 40mM, from about 30mM to about 35mM, from about 35mM to about 50mM, from about 35mM to about 45mM, from about 35mM to about 40mM, from about 40mM to about 50mM, from about 40mM to about 45mM, or from about 45mM to about 50mM). In some embodiments, the amount of dibasic sodium phosphate in the platelet additive solution is about 5mM, about lOmM, about l 5mM, about l6mM, about l7mM, about l8mM, about l9mM, about 20mM, about 2lmM, about 22mM, about 23mM, about 24mM, about 25mM, about 26mM, about 27mM, about 28mM, about 29mM, about 30mM, about 35mM, about 40mM, about 45mM, or about 50mM. In some embodiments, the amount of dibasic sodium phosphate in the platelet additive solution is about 2l .5mM or about 2l .6mM.

[0113] In some embodiments, the amount of monobasic sodium phosphate in the platelet additive solution is at least about lmM, at least about 5mM, at least about lOmM, at least about l5mM, at least about 20mM, at least about 25mM, at least about 30mM, at least about 35mM, at least about 40mM, or at least about 45mM. In some embodiments, the amount of dibasic sodium phosphate in the platelet additive solution is from about lmM to about 45mM or any range of values thereof (e.g., from about lmM to about 40mM, from about lmM to about 35mM, from about lmM to about 30mM, from about lmM to about 25mM, from about lmM to about 20mM, from about lmM to about !5mM, from about lmM to about lOmM, from about lmM to about 5mM, from about 5mM to about 45mM, from about 5mM to about 40mM, from about 5mM to about 35mM, from about 5mM to about 30mM, from about 5mM to about 25mM, from about 5mM to about 20mM, from about 5mM to about l5mM, from about 5mM to about lOmM, from about lOmM to about 45mM, from about lOmM to about 40mM, from about lOmM to about 35mM, from about lOmM to about 30mM, from about lOmM to about 25mM, from about lOmM to about 20mM, from about lOmM to about l5mM, from about l5mM to about 45mM, from about l5mM to about 40mM, from about l5mM to about 35mM, from about l5mM to about 30mM, from about l5mM to about 25mM, or from about l5mM to about 20mM). In some embodiments, the amount of dibasic sodium phosphate in the platelet additive solution is about lmM, about 2mM, about 3mM, about 4mM, about 5mM, about 6mM, about 7mM, about 8mM, about 9mM, about lOmM, about l lmM, about l2mM, about l3mM, about l4mM, about l5mM, about 20mM, about 25mM, about 30mM, about 35mM, about 40mM, or about 45mM. In some embodiments, the amount of monobasic sodium phosphate in the platelet additive solution is about 6.7mM.

[0114] In some embodiments, the amount of acetate in the platelet additive solution is at least about 5mM, at least about lOmM, at least about l5mM, at least about 20mM, at least about 25mM, at least about 30mM, at least about 35mM, at least about 40mM, at least about 45mM, or at least about 50mM. In some embodiments, the amount of acetate in the platelet additive solution is from about 5mM to about 50mM or any range of values thereof ( e.g ., from about 5mM to about 45mM, from about 5mM to about 40mM, from about 5mM to about 35mM, from about 5mM to about 30mM, from about 5mM to about 25mM, from about 5mM to about 20mM, from about 5mM to about l5mM, from about 5mM to about lOmM, from about lOmM to about 50mM, from about lOmM to about 45mM, from about lOmM to about 40mM, from about lOmM to about 35mM, from about lOmM to about 30mM, from about lOmM to about 25mM, from about lOmM to about 20mM, from about lOmM to about l5mM, from about l5mM to about 50mM, from about l5mM to about 45mM, from about l5mM to about 40mM, from about l5mM to about 35mM, from about l5mM to about 30mM, from about l5mM to about 25mM, from about l5mM to about 20mM, from about 20mM to about 50mM, from about 20mM to about 45mM, from about 20mM to about 40mM, from about 20mM to about 35mM, from about 20mM to about 30mM, from about 20mM to about 25mM, from about 25mM to about 50mM, from about 25mM to about 45mM, from about 25mM to about 40mM, from about 25mM to about 35mM, from about 25mM to about 30mM, from about 30mM to about 50mM, from about 30mM to about 45mM, from about 30mM to about 40mM, from about 30mM to about 35mM, from about 35mM to about 50mM, from about 35mM to about 45mM, from about 35mM to about 40mM, from about 40mM to about 50mM, from about 40mM to about 45mM, or from about 45mM to about 50mM). In some embodiments, the amount of acetate in the platelet additive solution is about 5mM, about lOmM, about l5mM, about 20mM, about 25mM, about 26mM, about 27mM, about 28mM, about

29mM, about 30mM, about 3 lmM, about 32mM, about 33mM, about 34mM, about

35mM, about 36mM, about 37mM, about 38mM, about 39mM, about 40mM, about

45mM, or about 50mM. In some embodiments, the amount of acetate in the platelet additive solution is about 32.5mM.

[0115] In some embodiments, the amount of sodium acetate in the platelet additive solution is at least about 5mM, at least about lOmM, at least about l5mM, at least about 20mM, at least about 25mM, at least about 30mM, at least about 35mM, at least about 40mM, at least about 45mM, or at least about 50mM. In some embodiments, the amount of sodium acetate in the platelet additive solution is from about 5mM to about 50mM or any range of values thereof ( e.g ., from about 5mM to about 45mM, from about 5mM to about 40mM, from about 5mM to about 35mM, from about 5mM to about 30mM, from about 5mM to about 25mM, from about 5mM to about 20mM, from about 5mM to about l5mM, from about 5mM to about lOmM, from about lOmM to about 50mM, from about lOmM to about 45mM, from about lOmM to about 40mM, from about lOmM to about 35mM, from about lOmM to about 30mM, from about lOmM to about 25mM, from about lOmM to about 20mM, from about lOmM to about l5mM, from about l5mM to about 50mM, from about l5mM to about 45mM, from about l5mM to about 40mM, from about l5mM to about 35mM, from about l5mM to about 30mM, from about l5mM to about 25mM, from about l5mM to about 20mM, from about 20mM to about 50mM, from about 20mM to about 45mM, from about 20mM to about 40mM, from about 20mM to about 35mM, from about 20mM to about 30mM, from about 20mM to about 25mM, from about 25mM to about 50mM, from about 25mM to about 45mM, from about 25mM to about 40mM, from about 25mM to about 35mM, from about 25mM to about 30mM, from about 30mM to about 50mM, from about 30mM to about 45mM, from about 30mM to about 40mM, from about 30mM to about 35mM, from about 35mM to about 50mM, from about 35mM to about 45mM, from about 35mM to about 40mM, from about 40mM to about 50mM, from about 40mM to about 45mM, or from about 45mM to about 50mM). In some embodiments, the amount of sodium acetate in the platelet additive solution is about 5mM, about lOmM, about l5mM, about 20mM, about 25mM, about 26mM, about 27mM, about 28mM, about 29mM, about 30mM, about 3 lmM, about 32mM, about 33mM, about 34mM, about 35mM, about 36mM, about 37mM, about 38mM, about 39mM, about 40mM, about 45mM, or about 50mM. In some embodiments, the amount of sodium acetate in the platelet additive solution is about 32.5mM.

[0116] In some embodiments, the amount of chloride in the platelet additive solution is at least about 40mM, at least about 50mM, at least about 60mM, at least about 65mM, at least about 70mM, at least about 75mM, at least about 80mM, at least about 85mM, at least about 90mM, or at least about lOOmM. In some embodiments, the amount of chloride in the platelet additive solution is from about 40mM to about lOOmM or any range of values thereof ( e.g ., from about 40mM to about 90mM, from about 40mM to about 85mM, from about 40mM to about 80mM, from about 40mM to about 75mM, from about 40mM to about 70mM, from about 40mM to about 65mM, from about 40mM to about 60mM, from about 40mM to about 50mM, from about 50mM to about lOOmM, from about 50mM to about 90mM, from about 50mM to about 85mM, from about 50mM to about 80mM, from about 50mM to about 75mM, from about 50mM to about 70mM, from about 50mM to about 65mM, from about 50mM to about 60mM, from about 60mM to about lOOmM, from about 60mM to about 90mM, from about 60mM to about 85mM, from about 60mM to about 80mM, from about 60mM to about 75mM, from about 60mM to about 70mM, from about 60mM to about 65mM, from about 65mM to about lOOmM, from about 65mM to about 90mM, from about 65mM to about 85mM, from about 65mM to about 80mM, from about 65mM to about 75mM, from about 65mM to about 70mM, from about 70mM to about lOOmM, from about 70mM to about 90mM, from about 70mM to about 85mM, from about 70mM to about 80mM, from about 70mM to about 75mM, from about 75mM to about lOOmM, from about 75mM to about 90mM, from about 75mM to about 85mM, from about 75mM to about 80mM, from about 80mM to about lOOmM, from about 80mM to about 90mM, from about 80mM to about 85mM, from about 85mM to about lOOmM, from about 85mM to about 90mM, or from about 90mM to about lOOmM). In some embodiments, the amount of chloride in the platelet additive solution is about 40mM, about 50mM, about 60mM, about 65mM, about 66mM, about 67mM, about 68mM, about 69mM, about 70mM, about 7lmM, about 72mM, about 73mM, about 74mM, about 75mM, about 76mM, about 77mM, about 78mM, about 79mM, about 80mM, about 85mM, about 90mM, or about lOOmM. In some embodiments, the amount of chloride in the platelet additive solution is about 77.3mM or about 75.9mM.

[0117] In some embodiments, the amount of sodium chloride in the platelet additive solution is at least about 40mM, at least about 50mM, at least about 60mM, at least about 65mM, at least about 70mM, at least about 75mM, at least about 80mM, at least about 85mM, at least about 90mM, or at least about lOOmM. In some embodiments, the amount of sodium chloride in the platelet additive solution is from about 40mM to about lOOmM or any range of values thereof ( e.g ., from about 40mM to about lOOmM, from about 40mM to about 90mM, from about 40mM to about 85mM, from about 40mM to about 80mM, from about 40mM to about 75mM, from about 40mM to about 70mM, from about 40mM to about 65mM, from about 40mM to about 60mM, from about 40mM to about 50mM, from about 50mM to about lOOmM, from about 50mM to about 90mM, from about 50mM to about 85mM, from about 50mM to about 80mM, from about 50mM to about 75mM, from about 50mM to about 70mM, from about 50mM to about 65mM, from about 50mM to about 60mM, from about 60mM to about lOOmM, from about 60mM to about 90mM, from about 60mM to about 85mM, from about 60mM to about 80mM, from about 60mM to about 75mM, from about 60mM to about 70mM, from about 60mM to about 65mM, from about 65mM to about lOOmM, from about 65mM to about 90mM, from about 65mM to about 85mM, from about 65mM to about 80mM, from about 65mM to about 75mM, from about 65mM to about 70mM, from about 70mM to about lOOmM, from about 70mM to about 90mM, from about 70mM to about 85mM, from about 70mM to about 80mM, from about 70mM to about 75mM, from about 75mM to about lOOmM, from about 75mM to about 90mM, from about 75mM to about 85mM, from about 75mM to about 80mM, from about 80mM to about lOOmM, from about 80mM to about 90mM, from about 80mM to about 85mM, from about 85mM to about lOOmM, from about 85mM to about 90mM, or from about 90mM to about lOOmM). In some embodiments, the amount of sodium chloride in the platelet additive solution is about 40mM, about 50mM, about 60mM, about 65mM, about 66mM, about 67mM, about 68mM, about 69mM, about 70mM, about 7lmM, about 72mM, about 73mM, about 74mM, about 75mM, about 76mM, about 77mM, about 78mM, about 79mM, about 80mM, about 85mM, about 90mM, or about lOOmM. In some embodiments, the amount of sodium chloride in the platelet additive solution is about 77.3mM or about 75.9mM.

[0118] In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about 20mM to about 40mM sodium phosphate, from about 20mM to about 40mM sodium acetate, and from about 65mM to about 85mM sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about 25mM to about 35mM sodium phosphate, from about 25mM to about 35mM sodium acetate, and from about 70mM to about 80mM sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of about 28mM sodium phosphate, about 33mM sodium acetate, and about 77mM sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of about 28.2mM sodium phosphate, about 32.5mM sodium acetate, and about 77.3mM sodium chloride.

[0119] In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about lOmM to about 30mM dibasic sodium phosphate ( e.g ., anhydrous dibasic sodium phosphate), from about lmM to about l5mM monobasic sodium phosphate, from about 20mM to about 40mM sodium acetate, and from about 65mM to about 85mM sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about l5mM to about 25mM dibasic sodium phosphate (e.g., anhydrous dibasic sodium phosphate), from about 5 to about lOmM monobasic sodium phosphate, from about 25mM to about 35mM sodium acetate, and from about 70mM to about 80mM sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of about 22mM dibasic sodium phosphate (e.g., anhydrous dibasic sodium phosphate), about 7mM monobasic sodium phosphate, about 33mM sodium acetate, and about 77mM sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of about 2l.5mM dibasic sodium phosphate (e.g., anhydrous dibasic sodium phosphate), about 6.7mM monobasic sodium phosphate, about 32.5mM sodium acetate, and about 77.3mM sodium chloride. [0120] In other embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about 5mM to about 40mM sodium phosphate (e.g., monobasic sodium phosphate, dibasic sodium phosphate, or a combination thereof), from about 20mM to about 50mM sodium acetate, and from about 50mM to about 90mM sodium chloride. In other embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about 20mM to about 30mM sodium phosphate (e.g., monobasic sodium phosphate, dibasic sodium phosphate, or a combination thereof), from about 25mM to about 40mM sodium acetate, and from about 70mM to about 80mM sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about 25mM to about 30mM sodium phosphate (e.g., monobasic sodium phosphate, dibasic sodium phosphate, or a combination thereof), from about 25mM to about 35mM sodium acetate, and from about 70mM to about 80mM sodium chloride.

[0121] In other embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about lmM to about l5mM monobasic sodium phosphate, from about lOmM to about 30mM dibasic sodium phosphate (e.g., anhydrous dibasic sodium phosphate), from about 20mM to about 50mM sodium acetate, and from about 50mM to about 90mM sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about 5mM to about lOmM monobasic sodium phosphate, from about l5mM to about 25mM dibasic sodium phosphate (e.g., anhydrous dibasic sodium phosphate), from about 20mM to about 40mM sodium acetate, and from about 60mM to about 80mM sodium chloride. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about 5mM to about lOmM monobasic sodium phosphate, from about 20mM to about 25mM dibasic sodium phosphate (e.g., anhydrous dibasic sodium phosphate), from about 30mM to about 35mM sodium acetate, and from about 70mM to about 80mM sodium chloride.

[0122] In other embodiments, the platelet additive solution further comprises, consists essentially of, or consists of potassium, magnesium, glucose, calcium, hispidin, zinc, or a combination thereof. In some embodiments, the platelet additive solution further comprises, consists essentially of, or consists of from about lmM to about lOmM potassium, from about O.lmM to about 5mM magnesium, from about lmM to about 20mM glucose, from about O.OlmM to about 4mM calcium, or a combination thereof. In some embodiments, the potassium is potassium chloride. In some embodiments, the magnesium is magnesium chloride.

[0123] In other embodiments, the platelet additive solution comprises, consists essentially of, or consists of sodium phosphate ( e.g ., monobasic sodium phosphate, dibasic sodium phosphate, or a combination thereof), sodium acetate, sodium chloride, potassium chloride, magnesium chloride, and glucose. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of sodium phosphate (e.g., monobasic sodium phosphate, dibasic sodium phosphate, or a combination thereof), sodium acetate, sodium chloride, potassium chloride, magnesium chloride, and glucose, wherein the solution does not contain citrate. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about lOmM to about 40mM sodium phosphate (e.g., monobasic sodium phosphate, dibasic sodium phosphate, or a combination thereof), from about 20mM to about 50mM sodium acetate, from about 50mM to about 90mM sodium chloride, from about lmM to about lOmM potassium chloride, from about O.lmM to about 5mM magnesium chloride, and from about lmM to about 20mM glucose. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about 20mM to about 30mM sodium phosphate (e.g., monobasic sodium phosphate, dibasic sodium phosphate, or a combination thereof), from about 20mM to about 40mM sodium acetate, from about 60mM to about 70mM sodium chloride, from about lmM to about 5mM potassium chloride, from about lmM to about 2mM magnesium chloride, and from about lOmM to about 20mM glucose.

[0124] In other embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about 5mM to about lOmM monobasic sodium phosphate, from about l5mM to about 25mM dibasic sodium phosphate (e.g., anhydrous dibasic sodium phosphate), from about lOmM to about 40mM sodium acetate, from about 50mM to about 90mM sodium chloride, from about lmM to about lOmM potassium chloride, from about O.lmM to about 5mM magnesium chloride, and from about lmM to about 20mM glucose. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of from about 5mM to about lOmM monobasic sodium phosphate, from about 20mM to about 25mM dibasic sodium phosphate (e.g., anhydrous dibasic sodium phosphate), from about 20mM to about 40mM sodium acetate, from about 60mM to about 80mM sodium chloride, from about lmM to about 5mM potassium chloride, from about lmM to about 5mM magnesium chloride, and from about lOmM to about 20mM glucose.

[0125] In other embodiments, the platelet additive solution comprises, consists essentially of, or consists of about 30mM sodium acetate, about 7mM monobasic sodium phosphate, about 22mM dibasic sodium phosphate (e.g., anhydrous dibasic sodium phosphate), 70mM sodium chloride, about 5mM potassium chloride, about 2mM magnesium chloride, and about l7mM glucose. In some embodiments, the platelet additive solution comprises, consists essentially of, or consists of about 30mM sodium acetate, about 6.7mM monobasic sodium phosphate, about 2l.6mM dibasic sodium phosphate (e.g., anhydrous dibasic sodium phosphate), about 69.4mM sodium chloride, about 5mM potassium chloride, about l.5mM magnesium chloride, and about l6.8mM glucose.

[0126] In some embodiments, a platelet additive solution of the invention does not contain citrate or is citrate-free.

[0127] In some embodiments, the platelet additive solution has a pH of at least about 6.

In some embodiments, the platelet additive solution has a pH of at least about 6.2. In some embodiments, the platelet additive solution has a pH of about 6 or about 6.2.

[0128] In some embodiments, the platelet additive solution preserves platelet function, activation, or a combination thereof. In some embodiments, the platelet additive solution does not cause spontaneous clotting of platelets. In some embodiments, the platelet additive solution results in one or more of the following when used to store platelets:

(i) decreased levels of reactive oxygen species,

(ii) decreased glucose utilization,

(iii) decreased lactate production,

(iv) decreased P-selectin expression,

(v) decreased apoptosis,

(vi) increased aggregation to collagen,

(vii) increased aggregation to TRAP6,

(viii) increased cell function, and

(ix) increased cell triggered activation.

[0129] In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride and citrate. [0130] In some embodiments, the platelet additive solution results in one or more of the following when used to store platelets:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation.

[0131] In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride and citrate.

[0132] In some embodiments, the platelet additive solution results in one or more of the following when used to store platelets:

(i) a decrease of from about 30% to about 60% in reactive oxygen species levels, or any values or range of values thereof (e.g., from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 60%, from about 40% to about 50%, about 30%, about 40%, about 50%, or about 60%),

(ii) a decrease of from about 30% to about 60% in cell death, or any values or range of values thereof (e.g., from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 60%, from about 40% to about 50%, about 30%, about 40%, about 50%, or about 60%),

(iii) a decrease of from about 40% to about 80% in glucose utilization, or any values or range of values thereof (e.g., from about 40% to about 70%, from about 40% to about 60%, from about 40% to about 50%, from about 50% to about 80%, from about 50% to about 70%, from about 50% to about 60%, from about 60% to about 80%, from about 60% to about 70%, from about 70% to about 80%, about 40%, about 50%, about 60%, about 70%, or about 80%),

(iv) a decrease of from about 30% to about 60% in lactate production, or any values or range of values thereof (e.g., from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 60%, from about 40% to about 50%, about 30%, about 40%, about 50%, or about 60%), (v) a decrease of from about 20% to about 50% in CD62P expression, or any values or range of values thereof (e.g., from about 20% to about 40%, from about 20% to about 30%, from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 50%, about 20%, about 30%, about 40%, or about 50%),

(vi) an increase of from about 10% to about 50% in collagen aggregation, or any values or range of values thereof (e.g., from about 10% to about 40%, from about 10% to about 30%, from about 10% to about 20%, from about 20% to about 50%, from about 20% to about 40%, from about 20% to about 30%, from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 50%, about 10%, about 20%, about 30%, about 40%, or about 50%), and

(vii) an increase of from about 10% to about 50% in TRAP6 aggregation, or any values or range of values thereof (e.g., from about 10% to about 40%, from about 10% to about 30%, from about 10% to about 20%, from about 20% to about 50%, from about 20% to about 40%, from about 20% to about 30%, from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 50%, about 10%, about 20%, about 30%, about 40%, or about 50%).

In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate. In some embodiments, the citrate in the reference platelet additive solution is at least lmM citrate. In some embodiments, the citrate in the reference platelet additive solution is from about lmM to about 20mM citrate or any values of range of values thereof (e.g., about lmM, about 5mM, about lOmM, about l5mM, or about 20mM). In some embodiments, the citrate in the reference platelet additive solution is sodium citrate. In some embodiments, the citrate in the reference platelet additive solution is at least lmM sodium citrate. In some embodiments, the citrate in the reference platelet additive solution is from about lmM to about 20mM sodium citrate or any values or range of values thereof (e.g., about lmM, about 5mM, about lOmM, about l5mM, or about 30mM).

[0133] In some embodiments, the platelet additive solution is for storage of platelets for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any range of values thereof (e.g., from about 1 day to about 10 days, from about 1 days to about 9 days, from about 1 day to about 8 days, from about 1 day to about 7 days, from about 1 day to about 6 days, from about 1 day to about 5 days, from about 1 day to about 4 days, from about 1 day to about 3 days, from about 1 day to about 2 days, from about 2 days to about 10 days, from about 2 days to about 9 days, from about 2 days to about 8 days, from about 2 days to about 7 days, from about 2 days to about 6 days, from about 2 days to about 5 days, from about 2 days to about 4 days, from about 2 days to about 3 days, from about 3 days to about 10 days, from about 3 days to about 9 days, from about 3 days to about 8 days, from about 3 days to about 7 days, from about 3 days to about 6 days, from about 3 days to about 5 days, from about 3 days to about 4 days, from about 4 days to about 10 days, from about 4 days to about 9 days, from about 4 days to about 8 days, from about 4 days to about 7 days, from about 4 days to about 6 days, from about 4 days to about 5 days, from about 5 days to about 10 days, from about 5 days to about 9 days, from about 5 days to about 8 days, from about 5 days to about 7 days, from about 5 days to about 6 days, from about 6 days to about 10 days, from about 6 days to about 9 days, from about 6 days to about 8 days, from about 6 days to about 7 days, from about 7 days to about 10 days, from about 7 days to about 9 days, from about 7 days to about 8 days, from about 8 days to about 10 days from about 8 days to about 9 days, or from about 9 days to about 10 days). In some embodiments, the platelet additive solution is for storage of platelets for about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or about 10 days.

[0134] In other embodiments, the platelet additive solution is for storage of platelets treated with a pathogen reduction system. In some embodiments, the pathogen reduction system is ultraviolet light exposure and/or a reagent that inhibits the replication of DNA or RNA. In some embodiments, the pathogen reduction system is a psoralen and/or ultraviolet A (UVA) light.

[0135] In other embodiments, the present invention relates to a composition comprising, consisting essentially of, or consisting of a platelet additive solution described herein and platelets. In some embodiments, the composition has a pH of at least about 6. In some embodiments, the composition has a pH of at least about 6.2. In some embodiments, the composition has a pH of about 6 or about 6.2.

[0136] In some embodiments, the composition preserves platelet function, activation, or a combination thereof. In some embodiments, the composition does not cause spontaneous clotting of platelets. In some embodiments, the composition results in one or more of the following:

(i) decreased levels of reactive oxygen species,

(ii) decreased glucose utilization,

(iii) decreased lactate production,

(iv) decreased P-selectin expression,

(v) decreased apoptosis,

(vi) increased aggregation to collagen,

(vii) increased aggregation to TRAP6,

(viii) increased cell function, and

(ix) increased cell triggered activation.

[0137] In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0138] In some embodiments, the composition results in one or more of the following:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation.

[0139] In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0140] In some embodiments, the composition results in one or more of the following:

(ii) a decrease of from about 30% to about 60% in cell death, or any values or range of values thereof (e.g., from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 60%, from about 40% to about 50%, about 30%, about 40%, about 50%, or about 60%), (iii) a decrease of from about 40% to about 80% in glucose utilization, or any values or range of values thereof (e.g., from about 40% to about 70%, from about 40% to about 60%, from about 40% to about 50%, from about 50% to about 80%, from about 50% to about 70%, from about 50% to about 60%, from about 60% to about 80%, from about 60% to about 70%, from about 70% to about 80%, about 40%, about 50%, about 60%, about 70%, or about 80%),

(iv) a decrease of from about 30% to about 60% in lactate production, or any values or range of values thereof (e.g., from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 60%, from about 40% to about 50%, about 30%, about 40%, about 50%, or about 60%),

(v) a decrease of from about 20% to about 50% in CD62P expression, or any values or range of values thereof (e.g., from about 20% to about 40%, from about 20% to about 30%, from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 50%, about 20%, about 30%, about 40%, or about 50%),

[0141] In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate. In some embodiments, the citrate in the reference platelet additive solution is at least lmM citrate. In some embodiments, the citrate in the reference platelet additive solution is from about lmM to about 20mM citrate or any values of range of values thereof (e.g., about lmM, about 5mM, about lOmM, about !5mM, or about 20mM). In some embodiments, the citrate used in the reference platelet additive solution is sodium citrate. In some embodiments, the citrate used in the reference platelet additive solution is at least lmM sodium citrate. In some embodiments, the citrate in the reference platelet additive solution is from about lmM to about 20mM sodium citrate or any values or range of values thereof (e.g., about lmM, about 5mM, about lOmM, about l5mM, or about 30mM).

[0142] In some embodiments, the composition stores platelets for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any range of values thereof (e.g., from about 1 day to about 10 days, from about 1 days to about 9 days, from about 1 day to about 8 days, from about 1 day to about 7 days, from about 1 day to about 6 days, from about 1 day to about 5 days, from about 1 day to about 4 days, from about 1 day to about 3 days, from about 1 day to about 2 days, from about 2 days to about 10 days, from about 2 days to about 9 days, from about 2 days to about 8 days, from about 2 days to about 7 days, from about 2 days to about 6 days, from about 2 days to about 5 days, from about 2 days to about 4 days, from about 2 days to about 3 days, from about 3 days to about 10 days, from about 3 days to about 9 days, from about 3 days to about 8 days, from about 3 days to about 7 days, from about 3 days to about 6 days, from about 3 days to about 5 days, from about 3 days to about 4 days, from about 4 days to about 10 days, from about 4 days to about 9 days, from about 4 days to about 8 days, from about 4 days to about 7 days, from about 4 days to about 6 days, from about 4 days to about 5 days, from about 5 days to about 10 days, from about 5 days to about 9 days, from about 5 days to about 8 days, from about 5 days to about 7 days, from about 5 days to about 6 days, from about 6 days to about 10 days, from about 6 days to about 9 days, from about 6 days to about 8 days, from about 6 days to about 7 days, from about 7 days to about 10 days, from about 7 days to about 9 days, from about 7 days to about 8 days, from about 8 days to about 10 days from about 8 days to about 9 days, or from about 9 days to about 10 days). In some embodiments, the composition stores platelets for about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or about 10 days.

[0143] In other embodiments, the platelets of the composition are treated with a pathogen reduction system. In some embodiments, the pathogen reduction system is ultraviolet light exposure and/or a reagent that inhibits the replication of DNA or RNA. In some embodiments, the pathogen reduction system is a psoralen and/or ultraviolet A (UVA) light. In some embodiments, the composition has at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 100% fewer pathogens. In some embodiments, the composition has from about 10% to about 100% fewer pathogens or any values or range of values thereof ( e.g ., from about 10% to about 80%, from about 10% to about 50%, from about 20% to about 100%, from about 20% to about 80%, or from about 20% to about 50%).

[0144] In some embodiments, platelets treated with a pathogen reduction system exhibit one of more of the following when combined with a platelet additive solution of the invention:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation, or any other results described herein with regard to the platelet additive solutions of the invention.

[0145] In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate (e.g., sodium citrate at an amount described herein).

Methods of Use and Making

[0146] In some embodiments, the present invention relates to a method for storing platelets comprising (i) combining platelets with a platelet additive solution described herein and (ii) storing the combination of (i). In some embodiments, the method comprises storing the combination of (i) for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any other values or ranges of values described herein.

[0147] In some embodiments, the present invention relates to a method for storing platelets comprising (i) providing a platelet composition (e.g., a composition comprising, consisting essentially of, or consisting of a platelet additive solution described herein and platelets) and (ii) storing the composition. In some embodiments, the method comprises storing the composition for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any other values or ranges of values described herein.

[0148] In some embodiments of the method for storing, the platelets exhibit one or more of the following after storage:

(i) decreased levels of reactive oxygen species,

(ii) decreased glucose utilization,

(iii) decreased lactate production,

(iv) decreased P-selectin expression,

(v) decreased apoptosis,

(vi) increased aggregation to collagen,

(vii) increased aggregation to TRAP6,

(viii) increased cell function, and

(ix) increased cell triggered activation.

[0149] In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0150] In some embodiments of the method for storing, the platelets exhibit one or more of the following after storage:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation.

[0151] In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0152] In some embodiments of the method for storing, the platelets exhibit one or more of the following after storage: (i) a decrease of from about 30% to about 60% in reactive oxygen species levels, or any values or range of values thereof (e.g., from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 60%, from about 40% to about 50%, about 30%, about 40%, about 50%, or about 60%),

[0153] In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0154] In some embodiments, the present invention relates to a method for increasing platelet function comprising (i) combining platelets with a platelet additive solution described herein and (ii) storing the combination of (i). In some embodiments, the method comprises storing the combination of (i) for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any other values or ranges of values described herein.

[0155] In some embodiments, the present invention relates to a method for increasing platelet function comprising (i) providing a platelet composition ( e.g ., a composition comprising, consisting essentially of, or consisting of a platelet additive solution described herein and platelets) and (ii) storing the composition. In some embodiments, the method comprises storing the composition for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any other values or ranges of values described herein.

[0156] In some embodiments of the method for increasing platelet function, the platelets have (i) a decrease of at least about 30% in reactive oxygen species levels, (ii) a decrease of at least about 40% in glucose utilization, (iii) a decrease of at least about 30% in lactate production, or (iv) a combination thereof, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0157] In some embodiments of the method for increasing platelet function, the platelets have:

(ii) a decrease of from about 40% to about 80% in glucose utilization, or any values or range of values thereof (e.g., from about 40% to about 70%, from about 40% to about 60%, from about 40% to about 50%, from about 50% to about 80%, from about 50% to about 70%, from about 50% to about 60%, from about 60% to about 80%, from about 60% to about 70%, from about 70% to about 80%, about 40%, about 50%, about 60%, about 70%, or about 80%),

(iii) a decrease of from about 30% to about 60% in lactate production, or any values or range of values thereof (e.g., from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 60%, from about 40% to about 50%, about 30%, about 40%, about 50%, or about 60%), or

(iv) a combination thereof, after storage.

[0158] In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0159] In some embodiments, the present invention relates to a method for decreasing platelet activation or aggregation during storage comprising (i) combining platelets with a platelet additive solution described herein and (ii) storing the combination of (i). In some embodiments, the method comprises storing the combination of (i) for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any other values or ranges of values described herein. In some embodiments, the present invention relates to a method for decreasing platelet activation or aggregation during storage comprising (i) providing a platelet composition (e.g., a composition comprising, consisting essentially of, or consisting of a platelet additive solution described herein and platelets) and (ii) storing the composition. In some embodiments, the method comprises storing the composition for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any other values or ranges of values described herein. In some embodiments of the method for decreasing platelet activation or aggregation during storage, the platelets have an increase of at least about 15% activation. In some embodiments of the method for decreasing platelet activation or aggregation during storage, the platelets have a decrease of at least about 20% in CD62P expression. In some embodiments of the method for decreasing platelet activation or aggregation during storage, the platelets have a decrease of from about 20% to about 50% in CD62P expression, or any values or range of values thereof (e.g., from about 20% to about 40%, from about 20% to about 30%, from about 30% to about 50%, from about 30% to about 40%, about 20%, about 30%, about 40%, or about 50%). In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0160] In some embodiments, the present invention relates to a method of increasing triggered platelet activation or aggregation comprising (i) combining platelets with a platelet additive solution described herein and (ii) storing the combination of (i). In some embodiments, the method comprises storing the combination of (i) for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any other values or ranges of values described herein.

[0161] In some embodiments, the present invention relates to a method of increasing triggered platelet activation or aggregation comprising (i) providing a platelet composition ( e.g ., a composition comprising, consisting essentially of, or consisting of a platelet additive solution described herein and platelets) and (ii) storing the composition. In some embodiments, the method of increasing triggered platelet activation or aggregation comprises storing the composition for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any other values or ranges of values described herein.

[0162] In some embodiments of the method for increasing triggered platelet activation or aggregation, the platelets have an increase of at least about 10% in activation or aggregation. In some embodiments of the method for increasing triggered platelet activation or aggregation, the platelets have an increase of at least about 10% in collagen aggregation, an increase of at least about 10% in TRAP6 aggregation, or a combination thereof. In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

[0163] In some embodiments of the methods described herein, the platelets are stored from about 1 day to about 10 days, from about 1 day to about 7 days, or from about 1 day to about 5 days. In some embodiments of the methods described herein, the platelets are stored about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or about 10 days.

[0164] In some embodiments of the methods described herein, the citrate in the reference platelet additive solution is at least lmM citrate. In some embodiments, the citrate in the reference platelet additive solution is from about lmM to about 20mM citrate or any values or ranges of values thereof ( e.g ., about lmM, about 5mM, about lOmM, about l5mM, or about 20mM). In some embodiments, the citrate in the reference platelet additive solution is sodium citrate. In some embodiments, the citrate in the reference platelet additive solution is at least lmM sodium citrate. In some embodiments, the citrate in the reference platelet additive solution is from about lmM to about 20mM sodium citrate or any values or ranges of values thereof (e.g., about lmM, about 5mM, about lOmM, about l5mM, or about 30mM).

[0165] In other embodiments, the present invention relates to a method of treating or preventing hemorrhage in a subject in need thereof, comprising administering a platelet composition (e.g., a composition comprising, consisting essentially of, or consisting of a platelet additive solution described herein and platelets) to the subject. In some embodiments, the hemorrhage is acute or severe hemorrhage. In some embodiments, the platelet composition comprises from about 2.5 x 10¹¹ to about 5.5 x 10¹¹ platelets, or any amount or range of amounts thereof (e.g., from about 3 x 10¹¹ to about 5.5 x 10¹¹, from about 3.5 x 10¹¹ to about 5.5 x 10¹¹, from about 4 x 10¹¹ to about 5.5 x 10¹¹, from about 4.5 x 10¹¹ to about 5.5 x 10¹¹, from about 5 x 10¹¹ to about 5.5 x 10¹¹, from about 2.5 x 10¹¹ to about 5 x 10¹¹, from about 3 x 10¹¹ to about 5 x 10¹¹, from about 3.5 x 10¹¹ to about 5 x 10¹¹, from about 4 x 10¹¹ to about 5 x 10¹¹, from about 4.5 x 10¹¹ to about 5 x 10¹¹, from about 2.5 x 10¹¹ to about 4.5 x 10¹¹, from about 3 x 10¹¹ to about 4.5 x 10¹¹, from about 3.5 x 10¹¹ to about 4.5 x 10¹¹, from about 4 x 10¹¹ to about 4.5 x 10¹¹, from about 2.5 x 10¹¹ to about 4 x 10¹¹, from about 3 x 10¹¹ to about 4 x 10¹¹, from about 3.5 x 10¹¹ to about 4 x 10¹¹, from about 2.5 x 10¹¹ to about 3.5 x 10¹¹, from about 3 x 10¹¹ to about 3.5 x 10¹¹, from about 2.5 x 10¹¹ to about 3 x 10¹¹, about 2.5 x 10¹¹, about 3 x 10¹¹, about 3.5 x 10¹¹, about 4 x 10¹¹, about 4.5 x 10¹¹, about 5 x 10¹¹, or about 5.5 x 10¹¹ platelets).

[0166] In other embodiments, the present invention relates to a method of treating or preventing low platelet count in a subject in need thereof, comprising administering a platelet composition (e.g., a composition comprising, consisting essentially of, or consisting of a platelet additive solution described herein and platelets) to the subject. In some embodiments, the subject is undergoing chemotherapy. In some embodiments, the platelet composition comprises from about 2.5 x 10¹¹ to about 5.5 x 10¹¹ platelets, or any amount or range of amounts thereof (e.g., from about 3 x 10¹¹ to about 5.5 x 10¹¹, from about 3.5 x 10¹¹ to about 5.5 x 10^{1 1}, from about 4 x 10¹¹ to about 5.5 x 10¹¹, from about 4.5 x 10¹¹ to about 5.5 x 10^{1 1}, from about 5 x 10¹¹ to about 5.5 x 10^{1 1}, from about 2.5 x 10¹¹ to about 5 x 10^{1 1}, from about 3 x 10¹¹ to about 5 x 10^{1 1}, from about 3.5 x 10¹¹ to about 5 x 10¹¹, from about 4 x 10¹¹ to about 5 x 10^{1 1}, from about 4.5 x 10¹¹ to about 5 x 10^{1 1}, from about 2.5 x 10¹¹ to about 4.5 x 10^{1 1}, from about 3 x 10¹¹ to about 4.5 x 10^{1 1}, from about 3.5 x 10¹¹ to about 4.5 x 10^{1 1}, from about 4 x 10¹¹ to about 4.5 x 10^{1 1}, from about 2.5 x 10¹¹ to about 4 x 10^{1 1}, from about 3 x 10¹¹ to about 4 x 10^{1 1}, from about 3.5 x 10¹¹ to about 4 x 10^{1 1}, from about 2.5 x 10¹¹ to about 3.5 x 10^{1 1}, from about 3 x 10¹¹ to about 3.5 x 10^{1 1}, from about 2.5 x 10¹¹ to about 3 x 10¹¹, about 2.5 x 10^{1 1}, about 3 x 10^{1 1}, about 3.5 x 10¹¹, about 4 x 10¹¹, about 4.5 x 10^{1 1}, about 5 x 10^{1 1}, or about 5.5 x 10¹¹ platelets).

[0167] In some embodiments of the methods described herein, the subject is an animal.

In other embodiments, the subject is a mammal. In other embodiments, the subject is a human.

[0168] In other embodiments, the present invention relates to a method for reducing pathogens in a platelet composition comprising (i) combining platelets with a platelet additive solution described herein and (ii) storing the combination of (i). In some embodiments, the method comprises storing the combination of (i) for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any other values or ranges of values described herein.

[0169] In other embodiments, the present invention relates to a method for reducing pathogens in a platelet composition comprising (i) providing a platelet composition (e.g., a composition comprising, consisting essentially of, or consisting of a platelet additive solution described herein and platelets) and (ii) storing the composition. In some embodiments, the method comprises storing the composition for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, or any other values or ranges of values described herein.

[0170] In some embodiments of this method, the pathogen reduction system is ultraviolet light exposure and/or a reagent that inhibits the replication of DNA or RNA. In some embodiments, the pathogen reduction system is a psoralen and/or ultraviolet A (UVA) light.

[0171] In some embodiments of this method, the platelet composition has at least about

10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, or at least about 100% fewer pathogens. In some embodiments, the platelet composition has from about 10% to about 100% fewer pathogens or any values or range of values thereof (e.g., from about 10% to about 80%, from about 10% to about 50%, from about 20% to about 100%, from about 20% to about 80%, or from about 20% to about 50%). In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate (e.g., sodium citrate at an amount described herein).

[0172] In some embodiments of this method, platelets treated with a pathogen reduction system exhibit one of more of the following when combined with a platelet additive solution of the invention:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation, or any other results described herein with regard to the platelet additive solutions of the invention. In some embodiments, the results are relative to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate (e.g., sodium citrate at an amount described herein).

[0173] In other embodiments, the present invention relates to a method of making a platelet additive solution comprising combining phosphate, acetate and chloride (at any amount or range of amounts described herein) to form a solution. In other embodiments, the present invention relates to a method of making a platelet additive solution comprising combining sodium phosphate, sodium acetate, sodium chloride, potassium chloride, magnesium chloride, and glucose (at any amount or range of amounts described herein) to form a solution. In some embodiments, the platelet additive solution does not contain citrate. In some embodiments, the phosphate is sodium phosphate. In some embodiments, the sodium phosphate is dibasic sodium phosphate ( e.g ., anhydrous dibasic sodium phosphate), monobasic sodium phosphate, or a combination thereof. In some embodiments, the acetate is sodium acetate. In some embodiments, the chloride is sodium chloride.

EXAMPLES

[0174] Reference is now made to the following examples, which together with the above descriptions illustrate some embodiments of the invention in a non-limiting fashion.

EXAMPLE 1

Methods and Materials

[0175] The following methods and materials were used for the experiments described in the examples.

Materials

[0176] Phosphate Buffered Saline (PBS) and distilled water (dH₂0) were obtained from

Gibco (Fisher, Pittsburgh, PA). InterSol platelet additive solution was purchased from Fenwal (Deerfield, IL). Citrate-free PAS-3 (sodium chloride 77.3mM, sodium acetate 32.5mM, dibasic sodium phosphate anhydrous 2l.5mM, monobasic sodium phosphate 6.7mM) and Tyrode's buffer (138 mM NaCl, 2.7 mM KC1, 2 mM MgCl₂, 0.42 mM NaH₂P0₄, 5 mM glucose, 10 mM HEPES) were prepared by the American Red Cross (Washington, DC) using Coming Inc. (Corning, NY) 0.2m filtration units and adjusted to a pH 7.25 and pH 7.40 respectively. Platelet aliquots were stored in Amicus storage containers (ref #X6R2350) purchased from Fresenius Kabli (Deerfield, IL). Ethylenediaminetetraacetic acid (EDTA) 0.1M was prepared by the American Red Cross (Washington, DC). Collagen ( 1 pg/mL) and adenosine diphosphate (ADP) 1 OmM were obtained from Chronolog Corp. (Havertown, PA). Thrombin Receptor Activator Peptide 6 (TRAP6) and human serum albumin (HSA) were purchased from Sigma-Aldrich, Inc. (St. Louis, MO). CD6l-PerCP, CD62P-PE, CD42b-FITC, Annexin V-PE, and Annexin V Binding Buffer were purchased from Becton Dickinson (Immunocytometry Systems, San Jose, CA). MitoProbe 5-(and -6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H₂DCFDA) was purchased from Molecular Probes (Life Technologies, Grand Island, NY).

Methods

Platelet Collection and Study Design - Single Unit

[0177] An Amicus separator was used to collect a single unit of apheresis platelets (PLT) from blood samples of consenting human donors that had not taken aspirin for at least 2 days. A yield of 4.0 x 10¹¹ platelets in 100 mL plasma along with 90 mL concurrent plasma was targeted. The unit was weighed and citrate-free PAS-3 was added to obtain a final platelet additive solution (PAS) concentration of 65%. Four new empty Amicus platelet storage bags (ref #X6R2350) were sealed in half (~4 inches from inlet) using the Clamco bag sealer (Cleveland, OH). The surface to volume ratio of a half bag was calculated to store 60mL of product. The bags were labeled 1 through 4. Bag #1 represented OmM citrate. Bag #2 represented 2mM citrate. Bag #3 represented 5mM citrate. Bag #4 represented lOmM citrate. The collection yielded -285 mL of platelets and PAS, and 60mL was transferred to each bag by syringe using a CBS needle-free spike. A stock of 1M sodium citrate was prepared and 120pL was added to bag #2 (2mM final), 300mB to bag #3 (5mM final), and 600mB to bag #4 (lOmM final). The units were then allowed to rest on the benchtop at 22°C between 1-4 hours then transferred to an agitator at 22-24°C. Sampling and testing from all bags occurred on days 1, 5 and 7. The study was repeated four times.

Platelet Collection and Study Design - Double Unit/Full Bag

[0178] A double unit of apheresis PLT was collected on an Amicus separator from consenting human donors free from aspirin for at least 5 days, with a target yield of 8.0 x 10¹¹ of platelets in 200 mL plasma along with 90 mL concurrent plasma. Two equivalent aliquots -100 mL were transferred into Amicus storage containers. One of two additive solutions, citrate-free PAS-3 (bag #1) or InterSol (bag #2), were immediately added to the PLT bags to obtain a PAS concentration of 65%. The units were allowed to rest on the benchtop at 22°C between 1-4 hours then placed on the agitator at 22-24°C. Platelet parameters were measured on days 0, 1, 5 and 7. The study was repeated six times.

PLT Count and Blood Gases

[0179] Platelet samples were obtained by syringe using a CBS needle-free spike under sterile technique (OriGen Biomedical, Austin TX). PLT counts were measured using a Sysmex XE 2100D hematology analyzer (Sysmex Corp., Lincolnshire, IL) and total PLT content was determined by multiplying PLT count by aliquot volume. Glucose and lactate concentrations, partial pressures of carbon dioxide (pC0₂) and oxygen (p0₂), and pH at 37°C were measured using a blood gas analyzer (Cobas b22l, Roche Diagnostics, Indianapolis, IN). Measurements were obtained on days 0, 1, 5 and 7. Bicarbonate concentrations were calculated from C0₂ and pH values.

Chronolog Aggregation. Extent of Shape Change (ESC) and Hypertonic Shock

Response (HSR)

[0180] Platelet aggregation was assessed using a Chrono-log light transmission aggregometer (Model 700, Chrono-Log Corp., Havertown, PA). Apheresis PLT (diluted in autologous fresh frozen plasma or Tyrode's buffer to 3xl0⁸/mL) were activated with the following agonists: collagen lOmg/mL, adenosine diphosphate (ADP) lOmM, thrombin receptor activation peptide (TRAP6) 20mM, or a combination of both collagen and ADP. Tests were run for 5 minutes and percent platelet aggregation was reported. The extent of shape change (ESC) and hypotonic stress response (HSR) were also measured turbometrically as described in Holme et al ., Transfusion 1998; 38:31-40.

Flow Cytometry

[0181] Platelet surface characteristics and activation markers were analyzed by flow cytometry using a FACSCalibur (BD Biosciences). Platelet samples were diluted in PBS with 0.1% serum albumin to 1 x 10⁶ platelets/mL and incubated with CD6l-PerCP/CD62- PE/CD42b-FITC at saturating concentrations for 15 minutes at 22°C. PLT were identified using forward and side scatter characteristics and anti-CD6l binding. Fluorescence Minus One Controls (FMO) were used for establishing fluorochrome staining boundaries. Phosphatidyl serine (PS) expression was determined with annexin V conjugated with phosphatidyethanolamine (PE) and using manufacturer's supplied buffer according to manufacturer's protocol (Becton-Dickinson, Immunocytometry Systems). PS expression was reported as percentage of annexin V-positive platelets. Reactive Oxygen Species (ROS) were measured using CM-H2DCFDA. Platelets at a concentration of 1 x 10⁶ platelets/mL were separately stained with 5pmol/L of CMH2DCFDA for 15 minutes at 37°C in the dark and then acquired. The fluorescence resulting from the oxidation of 5pmol/L of the intracellular probe CM-H₂DCFDA (Molecular Probes, ThermoFisher Scientific) was utilized to measure ROS generation and reported as mean fluorescent intensity (MFI).

EXAMPLE 2

Effects of Citrate in Platelet Additive Solutions - Citrate Dose Response Experiments

[0182] A study was performed to evaluate different effects of sodium citrate at concentrations of OmM, 2mM, 5mM and lOmM in the following platelet additive solution (PAS) test formulation:

Table 1 : PAS Test Formulation

[0183] PAS test formulation with OmM citrate is also referred to as "citrate-free PAS-3" in these Examples.

[0184] After preparation of the test formulations, the effect of the test formulations on platelets was measured on days 1, 5 and 7 using the methods described above in Example 1 for the following parameters: cell count, blood gas analysis, flow cytometry surface (CD42b), and activation markers (CM-H2DCFDA, AnnexinV and CD62P). Cell function was also analyzed by aggregometry to different agonists (TRAP6 and collagen).

Results

[0185] As shown in Table 2, platelet counts and blood gas profiles (bicarbonate, p0₂ and pC0₂ levels) were within acceptable limits for all platelet additive solutions tested and comparable throughout 7 days of platelet storage.

Table 2: Platelet count and blood gas profile. Results are from four separate experiments.

[0186] Next, reactive oxygen species levels following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or lOmM citrate were measured using flow cytometry and the CM-H2DCFDA probe, as described above in Example 1. FIG. 1A shows platelet additive solutions with lower or no citrate had significantly reduced reactive oxygen species levels. These results show that removal of citrate from platelet additive solutions significantly reduced oxidative stress.

[0187] Annexin V is a marker of cell death. Annexin V binding was measured following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or lOmM citrate using the methods described above in Example 1. FIG. 1B shows platelet additive solutions with lower amounts of citrate or no citrate had significantly reduced annexin V levels. These results show that removal of citrate from platelet additive solutions significantly reduced cell death.

[0188] Glucose utilization, lactate production, and pH were also measured following storage of platelets in test platelet additive solutions containing different amounts of citrate, or no citrate, using the methods described above in Example 1. FIG. 2 A shows glucose levels following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or lOmM citrate. These results show that removal of citrate from platelet additive solutions significantly preserved glucose levels and utilization.

[0189] FIG. 2B shows lactate levels following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or 10 mM citrate. These results show that removal of citrate from platelet additive solutions significantly reduced lactate levels.

[0190] FIG. 2C shows pH levels following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or 10 mM citrate. These results show that removal of citrate from platelet additive solution does not result in any significant change in pH over the course of 7 days.

[0191] Platelet surface markers were also measured following storage of platelets in test platelet additive solutions containing different amounts of citrate, or no citrate, using the methods described above in Example 1. FIG. 3 A shows levels of the platelet surface marker CD62P (P-selectin) following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or lOmM citrate. CD62P is an activation marker. As platelets become more and more activated, the levels of CD62P increase. These results show that removal of citrate significantly attenuated the level of CD62P at all time points.

[0192] FIG. 3B shows levels of the platelet surface receptor CD42b (GPIb) following storage of platelets for 1, 5 or 7 days in test platelet additive solutions containing OmM, 2mM, 5mM or lOmM citrate. CD42b expression levels have been shown to be reduced with platelet storage, and removal of this receptor from the platelet surface is thought to correlate with how long the platelet will circulate in the body following transfusion. As such, if CD42b expression levels remain high throughout storage, the platelets may be better for prophylactic platelet treatments. These results show a trend towards increased CD42b receptor expression levels in the test formulations that do not contain citrate.

EXAMPLE 3

Effects of Citrate in Platelet Additive Solutions - Aggregation

[0193] Platelet aggregation was measured following storage of platelets in test platelet additive solutions containing OmM or lOmM citrate using the methods described above in Example 1. Aggregations were performed in Tyrode's buffer or citrated plasma at a cell count of 300k/uL.

[0194] FIG. 4A shows aggregation data of platelets stimulated with collagen in Tyrode's buffer ("Tyrodes") and stored in test platelet additive solutions containing OmM or lOmM citrate for 1, 5 or 7 days (Dl, D5 and D7, respectively). FIG. 4B shows aggregation data of platelets stimulated with TRAP6 ("TRAP") in Tyrode's buffer ("Tyrodes") and stored in test platelet additive solutions containing OmM or lOmM citrate for 1, 5 or 7 days (Dl, D5 and D7, respectively). FIG. 4C shows aggregation data of platelets stimulated with collagen in plasma and stored in test platelet additive solutions containing OmM or lOmM citrate for 1, 5 or 7 days (Dl, D5 and D7, respectively). FIG. 4D shows aggregation data of platelets stimulated with TRAP6 ("TRAP") and in plasma and stored in test platelet additive solutions containing OmM or lOmM citrate for 1, 5 or 7 days (Dl, D5 and D7, respectively). Taken together, these data show significant improvements in platelet storage characteristics and platelet function when citrate is not included in the platelet additive solution.

EXAMPLE 4

Effects of Citrate in Platelet Additive Solutions - Full Bag Study Including Functional Data

Platelet Collection and Study Design

[0195] A double unit of apheresis PLT was collected on the Amicus separator as described above in Example 1. Various platelet parameters were measured on days 0, 1, 5, and 7. The study was repeated seven times.

Results

[0196] As shown in Table 3, platelet counts, blood gas profiles (bicarbonate, p0₂ and pC0₂ levels) and cell morphology were within acceptable limits for all platelet additive solutions tested and comparable throughout 7 days of platelet storage.

Table 3: Platelet count, blood gas profile, hypertonic shock response (HSR), and extent of shape change (ESR). Means and standard deviation (SD) are represented.

[0197] Next, reactive oxygen levels following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (OmM citrate) or InterSol (lOmM citrate) were measured using flow cytometry and the CM-H2DCFDA probe, as described above. FIG. 5A shows citrate-free PAS-3 had significantly reduced reactive oxygen levels. These results show that removal of citrate from the platelet additive solution significantly reduced oxidative stress.

[0198] Annexin V binding was also measured following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (OmM citrate) or InterSol (lOmM citrate) using the methods described above in Example 1. FIG. 5B shows citrate-free PAS-3 had significantly reduced annexin V levels. These results show that removal of citrate from the platelet additive solution significantly reduced cell death.

[0199] Glucose utilization, lactate production, and pH were also measured following storage of platelets in citrate-free PAS-3 or InterSol, using the methods described above in Example 1. FIG. 6A shows glucose levels following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (OmM citrate) or InterSol (lOmM citrate). These results show that removal of citrate from the platelet additive solution significantly preserved glucose levels and utilization. FIG. 6B shows lactate levels following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (OmM citrate) or InterSol (lOmM citrate). These results show that removal of citrate from the platelet additive solution significantly reduced lactate levels. FIG. 6C shows pH levels following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (OmM citrate) or InterSol (lOmM citrate). These results show that removal of citrate from the platelet additive solution does not result in any significant change in pH over the course of 7 days.

[0200] Platelet surface markers were also measured following storage of platelets in platelets in citrate-free PAS-3 or InterSol, using the methods described above in Example 1. FIG. 7A shows levels of the platelet surface marker CD62P (P-selectin) following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (OmM citrate) or InterSol (lOmM citrate). These results show that removal of citrate significantly attenuated the level of CD62P at all time points. FIG. 7B shows levels of the platelet surface receptor CD42b (GPIb) following storage of platelets for 1, 5 or 7 days in citrate-free PAS-3 (OmM citrate) or InterSol (lOmM citrate). These results show a trend towards increased CD42b receptor expression levels for test formulations that do not contain citrate.

[0201] Next, platelet aggregation was measured following storage of platelets in citrate- free PAS-3 (OmM citrate) or PAS-3 containing lOmM citrate in response to stimulation with collagen, TRAP6 or collagen and ADP on days 5 and 7 (except for day 7 collagen in plasma), using the methods described above in Example 1. FIGs. 8A, 8C and 8E show aggregation data for platelets resuspended in autologous plasma at a concentration of 300k/pL. FIGs. 8B, 8D and 8F show aggregation data for platelets resuspended in Tyrode's buffer ("Tyrodes"). Platelets in FIGs. 8A and 8B were stimulated with collagen. Platelets in FIGs. 8C and 8D were stimulated with TRAP6. Platelets in FIGs. 8E and 8F were stimulated with collagen and ADP. These data suggest the platelet microenvironment is important for overall platelet function and that citrated plasma inhibits the aggregation response. Platelets stored at room temperature in citrate-free PAS-3 had between an 8-19% increase in aggregation response to single agonists depending on the testing day and agonist used to trigger aggregation. FIGs. 8E and 8F show that citrate-free stored platelets respond to simultaneous stimulation with dual agonists and have a 13-15% increase in aggregation compared to those platelets stored in citrated PAS-3.

EXAMPLE 5

Effects of Citrate in Platelet Additive Solutions

[0202] A study was performed to evaluate the following platelet additive solution (PAS) test formulations: Table 4: PAS Test Formulations

[0203] After preparation of the test formulations, the effect of the test formulations on platelets was measured on days 0, 1, 5 and 7 for several different platelet characteristics, blood gas parameters and cell surface markers indicated in Table 5 below, using the methods described above in Example 1. Taken together, these data show significant improvement in several aspects when citrate is not included in a platelet additive solution.

Table 5: Blood Gas and Surface Marker Analysis

[0204] Next, platelet aggregation was also measured following storage of platelets in

PAS 5 A or PAS-P1 for 1, 5 or 8 days using the methods described above. Aggregations were performed at a cell count of 300k/uL. FIGs. 9A and 9C show aggregation data of platelets stimulated with collagen and stored in PAS 5 A or PAS-P1 for 1 day. Aggregation was evaluated in response to stimulation with collagen. FIGs. 9B and 9D show aggregation data of platelets stimulated with TRAP6 and stored in PAS 5 A or PAS- Pl for 1 day. FIGs. 10A and 10C show aggregation data of platelets stimulated with collagen and stored in PAS 5A or PAS-P1 for 5 days. FIGs. 10B and 10D show aggregation data of platelets stimulated with TRAP6 and stored in PAS 5 A or PAS-P1 for 5 days. FIGs. 11A and 11C show aggregation data of platelets stimulated with collagen and stored in PAS 5A or PAS-P1 for 8 days. FIGs. 11B and 11D show aggregation data of platelets stimulated with TRAP6 and stored in PAS 5A or PAS-P1 for 8 days. Taken together, these data show significant improvements in platelet storage characteristics and platelet function when citrate is not included in a platelet additive solution. EXAMPLE 6

Pathogen Reduction Study - A Psoralen and Ultraviolet A Light

Platelet Collection and Study Design

[0205] A double unit of apheresis PLT was collected on an Amicus separator from consenting human donors free from aspirin for at least 5 days, with a target yield of 8.0 x 10¹¹ cells in 200 mL plasma along with 90 mL concurrent plasma. Two equivalent aliquots of -100 mL were transferred into Amicus storage containers. One of two platelet additive solutions, citrate-free PAS-3 (Example 1 above; bag #1) or InterSol (Example 1 above; bag #2), were immediately added to the PLT bags to obtain a PAS concentration of 65%. The units were allowed to rest on the benchtop at 22°C between 1-4 hours. Once the product was resuspended, samples were taken for pre-illumination testing. Testing included cell counts, blood gas analysis, flow cytometry testing, and functional aggregometry. A psoralen photochemical was then added to obtain a concentration of 120-180mM. Product was then transferred to a storage bag intended for ultraviolet A (UVA) light treatment. The platelet unit was treated with -3.60 J/cm². All testing parameters were then immediately examined with UVA treatment. The platelet unit was then transferred to a new storage container with a device contained within to remove free psoralen. The unit was placed on an agitator for 4 hours at 22-24°C. The platelet unit was then transferred to a final storage container and returned to the agitator.

[0206] After preparation of the units, the effect of the test formulations on platelets was measured on days 0, 1, 2 and 4 for several different platelet characteristics, blood gas parameters and cell surface markers indicated FIG. 12, using the methods described above in Example 1. Taken together, these data show significant improvement in several platelet characteristics when citrate is not included in a platelet additive solution.

[0207] Next, platelet aggregation was measured following storage of platelets in citrate- free PAS-3 or InterSol using the methods described above at time points pre-illumination or prior to exposure to UVA treatment or post-illumination or after exposure to UVA treatment. Aggregations were performed at a cell count of 300k/uL.

[0208] FIG. 13 A shows aggregation data of platelets stimulated with collagen and stored in citrate-free PAS-3 or InterSol at day 0, pre-illumination or prior to exposure of UVA treatment. FIG. 13B shows aggregation data of platelets stimulated with TRAP6 and stored in citrate-free PAS-3 or InterSol at day 0, pre-illumination or prior to exposure of UVA treatment. FIG. 13C shows aggregation data of platelets stimulated with collagen and adenosine diphosphate (ADP) and stored in citrate-free PAS-3 or InterSol at day 0, pre-illumination or prior to exposure of UVA treatment.

[0209] FIG. 14A shows aggregation data of platelets stimulated with collagen and stored in citrate-free PAS-3 or InterSol day 0, post-illumination or after exposure to UVA treatment. FIG. 14B shows aggregation data of platelets stimulated with TRAP6 and stored in citrate-free PAS-3 or InterSol day 0, post-illumination or after exposure to UVA treatment. FIG. 14C shows aggregation data of platelets stimulated with collagen and ADP and stored in citrate-free PAS-3 or InterSol at day 0, post-illumination or after exposure to UVA treatment.

[0210] FIG. 15A shows aggregation data of platelets stimulated with collagen and stored in citrate-free PAS-3 or InterSol at day 1, post-illumination or after exposure to UVA treatment. FIG. 15B shows aggregation data of platelets stimulated with TRAP6 and stored in citrate-free PAS-3 or InterSol at day 1, post-illumination or after exposure to UVA treatment. FIG. 15C shows aggregation data of platelets stimulated with collagen and ADP and stored in citrate-free PAS-3 or InterSol at day 1, post-illumination or after exposure to UVA treatment.

[0211] FIG. 16A shows aggregation data of platelets stimulated with collagen and stored in citrate-free PAS-3 or InterSol at day 4, post-illumination or after exposure to UVA treatment. FIG. 16B shows aggregation data of platelets stimulated with TRAP6 and stored in citrate-free PAS-3 or InterSol at day 4, post-illumination or after exposure to UVA treatment. FIG. 16C shows aggregation data of platelets stimulated with collagen and ADP and stored in citrate-free PAS-3 or InterSol at day 4, post-illumination or after exposure to UVA treatment.

[0212] Taken together, these data show significant improvements in platelet aggregation, storage characteristics and function when citrate is not included in a platelet additive solution.

[0213] All publications, patents and patent applications mentioned in this application are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that section headings are used, they should not be construed as necessarily limiting.

Claims

WHAT IS CLAIMED IS:

1. A platelet additive solution, comprising phosphate, acetate and chloride, wherein the solution does not contain citrate.

2. The solution of claim 1, wherein the phosphate is sodium phosphate.

3. The solution of claim 2, wherein the sodium phosphate is monobasic sodium phosphate.

4. The solution of claim 2, wherein the sodium phosphate is dibasic sodium phosphate.

5. The solution of claim 2, wherein the sodium phosphate is a combination of monobasic sodium phosphate and dibasic sodium phosphate.

6. The solution of any one of claims 1 to 5, wherein the acetate is sodium acetate.

7. The solution of any one of claims 1 to 6, wherein the chloride is sodium chloride.

8. A platelet additive solution, comprising sodium phosphate, sodium acetate, and sodium chloride, wherein the solution does not contain citrate.

9. The solution of claim 8, comprising from about 5mM to about 40mM sodium phosphate, from about 20mM to about 50mM sodium acetate, and from about 50mM to about 90mM sodium chloride.

10. The solution of claim 9, comprising from about 20mM to about 30mM sodium phosphate, from about 25mM to about 40mM sodium acetate, and from about 70mM to about 80mM sodium chloride.

11. The solution of claim 10, comprising from about 25mM to about 30mM sodium phosphate, from about 25mM to about 35mM sodium acetate, and from about 70mM to about 80mM sodium chloride.

12. A platelet additive solution, comprising monobasic sodium phosphate, dibasic sodium phosphate, sodium acetate, and sodium chloride, wherein the solution does not contain citrate.

13. The solution of claim 12, wherein the dibasic sodium phosphate is anhydrous dibasic sodium phosphate.

14. The solution of claim 12, comprising from about lmM to about l5mM monobasic sodium phosphate, from about lOmM to about 30mM dibasic sodium phosphate, from about 20mM to about 50mM sodium acetate, and from about 50mM to about 90mM sodium chloride.

15. The solution of claim 14, comprising from about 5mM to about lOmM monobasic sodium phosphate, from about l5mM to about 25mM dibasic sodium phosphate, from about 20mM to about 40mM sodium acetate, and from about 60mM to about 80mM sodium chloride.

16. The solution of claim 15, comprising from about 5mM to about lOmM monobasic sodium phosphate, from about 20mM to about 25mM dibasic sodium phosphate, from about 30mM to about 35mM sodium acetate, and from about 70mM to about 80mM sodium chloride.

17. The solution of any one of claims 1 to 16, further comprising potassium, magnesium, glucose, calcium, hispidin, zinc, or a combination thereof.

18. The solution of any one of claims 1 to 17, further comprising from about lmM to about lOmM potassium, from about O. lmM to about 5mM magnesium, from about lmM to about 20mM glucose, from about O.OlmM to about 4mM calcium, or a combination thereof.

19. A platelet additive solution, comprising sodium phosphate, sodium acetate, sodium chloride, potassium chloride, magnesium chloride and glucose, wherein the solution does not contain citrate.

20. The solution of claim 19, comprising from about lOmM to about 40mM sodium phosphate, from about 20mM to about 50mM sodium acetate, from about 50mM to about 90mM sodium chloride, from about lmM to about lOmM potassium chloride, from about O.lmM to about 5mM magnesium chloride, and from about lmM to about 20mM glucose.

21. The solution of claim 20, comprising from about 20mM to about 30mM sodium phosphate, from about 20mM to about 40mM sodium acetate, from about 60mM to about 70mM sodium chloride, from about lmM to about 5mM potassium chloride, from about lmM to about 2mM magnesium chloride, and from about lOmM to about 20mM glucose.

22. The solution of claim 19, comprising from about 5mM to about lOmM monobasic sodium phosphate, from about l5mM to about 25mM dibasic sodium phosphate, from about lOmM to about 40mM sodium acetate, from about 50mM to about 90mM sodium chloride, from about lmM to about lOmM potassium chloride, from about O. lmM to about 5mM magnesium chloride, and from about lmM to about 20mM glucose.

23. The solution of claim 22, wherein the dibasic sodium phosphate is anhydrous dibasic sodium phosphate.

24. The solution of claim 22, comprising from about 5mM to about lOmM monobasic sodium phosphate, from about 20mM to about 25mM dibasic sodium phosphate, from about 20mM to about 40mM sodium acetate, from about 60mM to about 80mM sodium chloride, from about lmM to about 5mM potassium chloride, from about lmM to about 5mM magnesium chloride, and from about lOmM to about 20mM glucose.

25. The solution of any one of claims 1 to 24 having a pH of about 6.2 or greater.

26. The solution of any one of claims 1 to 25, wherein the solution preserves platelet function, activation, or a combination thereof.

27. The solution of any one of claims 1 to 26, wherein the solution does not cause spontaneous clotting of platelets.

28. The solution of any one of claims 1 to 27, wherein the solution results in one or more of the following when used to store platelets:

(i) decreased levels of reactive oxygen species,

(ii) decreased glucose utilization,

(iii) decreased lactate production,

(iv) decreased P-selectin expression,

(v) decreased apoptosis,

(vi) increased aggregation to collagen,

(vii) increased aggregation to TRAP6,

(viii) increased cell function, and

(ix) increased cell triggered activation,

29. The solution of any one of claims 1 to 28, wherein the solution results in one or more of the following when used to store platelets:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation,

30. The solution of any one of claims 1 to 29, wherein the solution results in one or more of the following when used to store platelets:

(ii) a decrease of from about 30% to about 60% in cell death,

(iii) a decrease of from about 40% to about 80% in glucose utilization,

(iv) a decrease of from about 30% to about 60% in lactate production, (v) a decrease of from about 20% to about 50% in CD62P expression,

(vi) an increase of from about 10% to about 50% in collagen aggregation, and

(vii) an increase of from about 10% to about 50% in TRAP6 aggregation,

31. The solution of any one of claims 28 to 30, wherein the citrate in the reference platelet additive solution is from about lmM citrate to about 20mM citrate.

32. The solution of any one of claims 1 to 31, for storage of platelets for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, or at least 10 days.

33. The solution of any one of claims 1 to 32, for storage of platelets from about 1 day to about 10 days, from about 1 day to about 7 days, or from about 1 day to about 5 days.

34. The solution of any one of claims 1 to 33, for storage of platelets about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or about 10 days.

35. A composition, comprising (i) the platelet additive solution of any one of claims 1 to 34, and (ii) platelets.

36. A method for storing platelets, comprising (i) combining platelets with the platelet additive solution of any one of claims 1 to 34 or providing the composition of claim 35, and (ii) storing the combination or composition of (i) for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, or at least 10 days.

37. The method of claim 36, wherein the platelets exhibit one or more of the following after storage:

(i) decreased levels of reactive oxygen species,

(ii) decreased glucose utilization, (iii) decreased lactate production,

(iv) decreased P-selectin expression,

(v) decreased apoptosis,

(vi) increased aggregation to collagen,

(vii) increased aggregation to TRAP6,

(viii) increased cell function, and

(ix) increased cell triggered activation,

38. The method of claim 36 or 37, wherein the platelets exhibit one or more of the following after storage:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation,

39. The method of any one of claims 36 to 38, wherein the platelets exhibit one or more of the following after storage:

(ii) a decrease of from about 30% to about 60% in cell death,

(iii) a decrease of from about 40% to about 80% in glucose utilization,

(iv) a decrease of from about 30% to about 60% in lactate production,

(v) a decrease of from about 20% to about 50% in CD62P expression,

(vi) an increase of from about 10% to about 50% in collagen aggregation, and

(vii) an increase of from about 10% to about 50% in TRAP6 aggregation, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

40. A method for decreasing platelet activation or aggregation during storage, comprising (i) combining platelets with the platelet additive solution of any one of claims 1 to 34 or providing the composition of claim 35, and (ii) storing the combination or composition of (i) for least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, or at least 10 days.

41. The method of claim 40, wherein the platelets have (i) a decrease of at least about 30% in reactive oxygen species levels, (ii) a decrease of at least about 40% in glucose utilization, (iii) a decrease of at least about 30% in lactate production, (iv) a decrease of at least about 20% in CD62P expression, or (v) a combination thereof, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

42. The method of claim 40 or 41, wherein the platelets have (i) a decrease of from about 30% to about 60% in reactive oxygen species levels, (ii) a decrease of from about 40% to about 80% in glucose utilization, (iii) a decrease of from about 30% to about 60% in lactate production, (iv) a decrease of from about 20% to about 50% in CD62P expression, or (v) a combination thereof, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

43. A method for increasing triggered platelet activation or aggregation, comprising (i) combining platelets with the platelet additive solution of any one of claims 1 to 34 or the composition of claim 35, and (ii) storing the combination or composition of (i) for least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, or at least 10 days.

44. The method of claim 43, wherein the platelets have an increase of at least about 10% activation or aggregation, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

45. The method of claim 43 or 44, wherein the platelets have (i) a decrease of at least about 20% in CD62P expression, (ii) an increase of at least about 10% in collagen aggregation, (iii) an increase of at least about 10% in TRAP6 aggregation, or (iv) a combination thereof, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

46. The method of any one of claims 43 to 45, wherein the platelets have (i) a decrease of from about 20% to about 50% in CD62P expression, (ii) an increase of from about 10% to about 50% in collagen aggregation, (iii) an increase of from about 10% to about 50% in TRAP6 aggregation, or (iv) a combination thereof, after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

47. The method of any one of claims 36 to 46, wherein the platelets are stored from about 1 day to about 10 days, from about 1 day to about 7 days, or from about 1 day to about 5 days.

48. The method of any one of claims 36 to 47, wherein the platelets are stored about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, or about 10 days.

49. The method of any one of claims 36 to 48, wherein the citrate in the reference platelet additive solution is from about lmM citrate to about 20mM citrate.

50. A method of making a platelet additive solution, comprising combining phosphate, acetate and chloride to form a solution, wherein the solution does not contain citrate.

51. The method of claim 50, where the phosphate is sodium phosphate, the acetate is sodium acetate, and the chloride is sodium chloride.

52. The method of claim 51, wherein the sodium phosphate is a combination of monobasic sodium phosphate and dibasic sodium phosphate.

53. The method of claim 52, wherein the dibasic sodium phosphate is anhydrous dibasic sodium phosphate.

54. A method of making a platelet additive solution, comprising combining sodium phosphate, sodium acetate, sodium chloride, potassium chloride, magnesium chloride, and glucose to form a solution, wherein the solution does not contain citrate.

55. The method of claim 54, wherein the sodium phosphate is a combination of monobasic sodium phosphate and dibasic sodium phosphate.

56. A method of treating or preventing hemorrhage in a subject in need thereof, comprising administering the composition of claim 35 to the subject.

57. The method of claim 56, wherein the hemorrhage is acute or severe hemorrhage.

58. A method of treating or preventing low platelet count in a subject in need thereof, comprising administering the composition of claim 35 to the subject.

59. The method of claim 58, wherein the subject is undergoing chemotherapy.

60. The method of any one of claims 56 to 59, wherein the subject is a human.

61. The solution of any one of claims 26 to 34, wherein the platelets are treated with a pathogen reduction system.

62. The solution of claim 61, wherein the pathogen reduction system is ultraviolet light exposure and/or a reagent that inhibits the replication of DNA or RNA.

63. The composition of claim 35, wherein the platelets are treated with a pathogen reduction system.

64. The composition of claim 63, wherein the pathogen reduction system is ultraviolet light exposure and/or a reagent that inhibits the replication of DNA or RNA.

65. The method of any one of claims 36 to 60, wherein the platelets are treated with a pathogen reduction system.

66. The method of claim 65, wherein the pathogen reduction system is ultraviolet light exposure and/or a reagent that inhibits the replication of DNA or RNA.

67. A method for reducing pathogens in a platelet composition, comprising (i) combining platelets with the platelet additive solution of claim 61 or 62 or providing the composition of claim 63 or 64, and (ii) storing the combination or composition of (i) for least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 6 days, at least 9 days, or at least 10 days.

68. The method of claim 67, wherein the platelet composition has at least 10% fewer pathogens after storage, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.

69. A method for decreasing platelet activation or aggregation during storage or increasing triggered platelet activation or aggregation, comprising (i) combining platelets with the platelet additive solution of claim 61 or 62 or providing the composition of claim 63 or 64, and (ii) storing the combination or composition of (i) for least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 6 days, at least 9 days, or at least 10 days.

70. The method of claim 69, wherein the platelets exhibit one of more of the following after storage:

(i) a decrease of at least about 30% in reactive oxygen species levels,

(ii) a decrease of at least about 30% in cell death,

(iii) a decrease of at least about 40% in glucose utilization,

(iv) a decrease of at least about 30% in lactate production,

(v) a decrease of at least about 20% in CD62P expression,

(vi) an increase of at least about 10% in collagen aggregation, and

(vii) an increase of at least about 10% in TRAP6 aggregation, compared to a reference platelet additive solution comprising phosphate, acetate, chloride, and citrate.