Optimizing platelet-based therapeutic products through novel donor selection and pooling techniques

Abstract

This disclosure relates to therapeutic products, and includes methods for producing a consistent platelet-based therapeutic product by selecting donors based on donor's age, blood markers of inflammation, and specific cytokine and growth factor profiles and pooling their platelet concentrates. The method is useful for ensuring production of a consistent drug substance suitable for treating conditions such as osteoarthritis by reducing variability in the quality of the platelets, ensuring the platelets contain consistent concentrations of potentially beneficial growth factors and cytokines, and avoiding platelets with potentially age-related and inflammation-induced changes.

Description

Mintz Docket No.062397-501001WO PCT APPLICATION OPTIMIZING PLATELET-BASED THERAPEUTIC PRODUCTS THROUGH NOVEL DONOR SELECTION AND POOLING TECHNIQUES Inventors: Terence S. Russell, Residing at Irving, TX Assignee: Spry Bio Inc. 511 E John Carpenter Fwy Ste 500 Irving TX 75062-8138 Entity status: Small MINTZ LEVIN COHN FERRIS GLOVSKY AND POPEO, P.C. One Financial Center Boston, MA 02111 (617) 542-6000 Filed: October 16, 2025Mintz Docket No.062397-501001WO OPTIMIZING PLATELET-BASED THERAPEUTIC PRODUCTS THROUGH NOVEL DONOR SELECTION AND POOLING TECHNIQUES CROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 709,183, filedOctober 18, 2024, which application is incorporated herein by reference. BACKGROUND

[0002] There is a need for improved therapeutic products such as for treatment of osteoarthritis(OA). As of 2019, the global prevalence of osteoarthritis was estimated to affect approximately 528 million people. This number has been steadily increasing, with projections suggesting that by 2050, nearly 1 billion people will be living with osteoarthritis due to factors such as aging populations, population growth, and rising obesity rates. In the United States alone, the prevalence of osteoarthritis in 2019 was estimated to be about 58.5 million adults. This figure represents a significant portion of the adult population, reflecting the high burden of the disease within the U.S. context.

[0003] These figures underscore the significant and growing burden of osteoarthritis bothglobally and within the United States. The increase in prevalence over the past decades highlights the urgent need for effective treatments and interventions to manage and potentially mitigate the impact of OA on populations worldwide. SUMMARY

[0004] Provided herein, in some embodiments, are methods for producing a consistent andoptimized platelet-based therapeutic product. This may be accomplished through donor selection and pooling of platelet concentrates. Donor selection may include selecting platelet donors based on (1) age and inflammatory status to reduce negative age-related and inflammation-related changes in platelets, and (2) using scientifically predefined cytokine and growth factor profiles that are critical for having consistent therapeutic potential in the drug substance and ensuring its suitability for treating osteoarthritis and related conditions. Pooling of platelet concentrates may include pooling from selected donors to create a standardized therapeutic product with reduced variability in key biological factors, thereby enhancing the consistency of the final drug substance.

[0005] Provided herein, in some embodiments, are methods of making a platelet composition.Some embodiments of the method include: obtaining donor information for a group of human donors; optionally obtaining biofluid samples of the group of human donors; obtaining platelet-enriched samples of the group of human donors; and for a donor of the group of donors: determining an age of the donor from the donor information, determining a disease status of the donor from the donor information or from disease status measurements obtained from one or more of the biofluid samples, obtaining a systemic inflammation measurement for the donor from the donor information or from aMintz Docket No.062397-501001WO systemic inflammation measurement obtained from one or more of the biofluid samples, obtaining cytokine or growth factor measurements for the donor from a platelet-enriched sample of the donor, pooling the platelet-enriched sample of the donor with other platelet-enriched samples when the donor age is within a predetermined age range, the donor disease status is free of a predetermined selection of diseases, the systemic inflammation measurement is below a predetermined systemic inflammation threshold, and the cytokine or growth factor measurements are within predetermined cytokine or growth factor ranges, thereby generating a platelet composition comprising pooled platelet-enriched samples of the group of human donors, and not pooling the platelet-enriched sample of the donor with the other platelet-enriched samples when the donor age is not within the predetermined age range, the donor disease status is not free of the of diseases, the systemic inflammation measurement is not below the predetermined systemic inflammation threshold, or the cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges. In some embodiments, the platelet-enriched samples are generated via a series of centrifugation steps or by apheresis of a blood sample. In some embodiments, the biofluid sample is a whole blood sample or a plasma sample. In some embodiments, obtaining the platelet-enriched sample of the donor is optional, and wherein: the platelet-enriched sample of the donor is obtained when the donor age is within the predetermined age range, the donor disease status is free of the predetermined selection of diseases, and the systemic inflammation measurement is below the predetermined systemic inflammation threshold; and the platelet-enriched sample of the donor is not obtained when the donor age is not within the predetermined age range, the donor disease status is not free the predetermined selection of diseases, or the systemic inflammation measurement is not below the predetermined systemic inflammation threshold, wherein when the platelet-enriched sample of the donor is not obtained, no platelet-enriched sample of the donor is pooled with the other platelet-enriched samples. In some embodiments, the predetermined age range includes an age of at least 10 years, at least 12 years, at least 14 years, at least 16 years, at least 18 years, at least 20 years, at least 22 years, at least 24 years, at least 26 years, at least 28 years, or at least 30 years. In some embodiments, the predetermined age range includes an age below 60 years, below 58 years, below 56 years, below 54 years, below 52 years, below 50 years, below 48 years, below 46 years, below 44 years, below 42 years, below 40 years, below 38 years, below 36 years, below 34 years, or below 32 years, or below 30 years. In some embodiments, the predetermined selection of diseases comprises infectious disease, cancer, autoimmune disease, and cardiovascular disease. In some embodiments, the predetermined selection of diseases comprises human immunodeficiency virus, human T-cell lymphotropic virus, hepatitis B, hepatitis C, syphilis, West Nile virus, and Chagas disease. In some embodiments, the systemic inflammation measurement comprises a C-reactive protein (CRP) measurement, and the predetermined systemic inflammation threshold comprises a predetermined CRP threshold. In some embodiments, the predetermined CRP threshold is 0 mg / L (e.g. undetectable), 0.5 mg / L, 1 mg / L, 1.5 mg / L, 2 mg / L, 2.5 mg / L, 3 mg / L, 3.5 mg / L, 4 mg / L, 4.5 mg / L, 5 mg / L, 5.5 mg / L, 6 mg / L, 6.5 mg / L,Mintz Docket No.062397-501001WO 7 mg / L, 7.5 mg / L, 8 mg / L, 8.5 mg / L, 9 mg / L, 9.5 mg / L, or 10 mg / L. In some embodiments, the systemic inflammation measurement comprises an erythrocyte sedimentation rate (ESR) measurement, and the predetermined systemic inflammation threshold comprises a predetermined ESR threshold. In some embodiments, the predetermined ESR threshold is 10 mm / hour, 15 mm / hour, 20 mm / hour, 25 mm / hour, 30 mm / hour, 35 mm / hour, or 40 mm / hour. In some embodiments, the predetermined cytokine or growth factor ranges are within 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, or 3 standard deviations of average cytokine or growth factor measurements in a normal population. In some embodiments, a predetermined cytokine or growth factor range of the predetermined cytokine or growth factor ranges is at least 50 pg / mL, at least 100 pg / mL, at least 500 pg / mL, at least 1 µg / mL, at least 5 µg / mL, at least 10 µg / mL, at least 50 µg / mL, or at least 100 µg / mL. In some embodiments, a predetermined cytokine or growth factor range of the predetermined cytokine or growth factor ranges is below 100 pg / mL, below 500 pg / mL, below 1 µg / mL, below 5 µg / mL, below 10 µg / mL, below 50 µg / mL, below 100 µg / mL, or below 500 µg / mL. In some embodiments, the predetermined cytokine or growth factor ranges comprise ranges for each of a plurality of cytokine(s) and / or growth factor(s). In some embodiments, the cytokine or growth factor measurements are combined into a cytokine or growth factor index. In some embodiments, combining the cytokine or growth factor measurements into a cytokine or growth factor index comprises obtaining an arithmetic mean, a geometric mean, a median, or a weighted average. In some embodiments, the predetermined cytokine or growth factor ranges comprise a predetermined cytokine or growth factor index range. In some embodiments, the cytokine or growth factor measurements comprise a chemokine C-C motif ligand 2 (CCL-2) measurement. In some embodiments, the cytokine or growth factor measurements comprise a connective tissue growth factor (CTGF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a fibroblast growth factor (FGF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a fibroblast growth factor 2 (FGF-2) measurement. In some embodiments, the cytokine or growth factor measurements comprise a hepatocyte growth factor (HGF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a histamine measurement. In some embodiments, the cytokine or growth factor measurements comprise an insulin-like growth factor 1 (IGF-1) measurement. In some embodiments, the cytokine or growth factor measurements comprise aninterleukin-1 beta (IL-1β) measurement. In some embodiments, the cytokine or growth factormeasurements comprise a platelet-derived growth factor (PDGF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a platelet-derived growth factor BB (PDGF-BB) measurement. In some embodiments, the cytokine or growth factor measurements comprise a platelet factor 4 (PF4) measurement. In some embodiments, the cytokine or growth factor measurements comprise a prostaglandin E2 measurement. In some embodiments, the cytokine orgrowth factor measurements comprise a transforming growth factor-beta (TGF-β) measurement. Insome embodiments, the cytokine or growth factor measurements comprise a transforming growthMintz Docket No.062397-501001WOfactor-beta 1 (TGF-β1) measurement. In some embodiments, the cytokine or growth factormeasurements comprise a tumor necrosis factor (TNF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a thrombospondin-1 (TSP-1) measurement. In some embodiments, the cytokine or growth factor measurements comprise an epidermal growth factor (EGF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a vascular endothelial growth factor (VEGF) measurement. Some embodiments include enriching the platelet composition further by centrifuging the pooled platelet-enriched samples and removing erythrocytes and leukocytes. Some embodiments include lyophilizing the platelet composition. Some embodiments include obtaining second cytokine or growth factor measurements from the platelet composition before the lyophilization. Some embodiments include lyophilizing the platelet composition when the second cytokine or growth factor measurements are within the predetermined cytokine or growth factor ranges, and not lyophilizing the platelet composition when the second cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges. Some embodiments include reconstituting the lyophilized platelet composition. Some embodiments include combining the platelet composition with a buffer. Some embodiments include the platelet composition with a monosaccharide, disaccharide, or polysaccharide. Some embodiments include formulating the platelet composition into a pharmaceutical composition. In some embodiments, the pharmaceutical composition comprises a liquid solution. In some embodiments, the pharmaceutical composition comprises a lyophilized platelet composition. In some embodiments, the lyophilized platelet composition is formulated for reconstitution with a reconstitution buffer by a medical practitioner. Some embodiments include packaging the platelet composition. Some embodiments include obtaining second or third cytokine or growth factor measurements from the platelet composition before the packaging. Some embodiments include packaging the platelet composition when the second or third cytokine or growth factor measurements are within the predetermined cytokine or growth factor ranges, and not packaging the platelet composition when the second or third cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges. Some embodiments include administering the platelet composition to a subject in need of treatment for a disease or disorder. In some embodiments, the disease comprises an autoimmune disease, an inflammatory disease, a degenerative joint disease. In some embodiments, the disease or disorder comprises osteoarthritis, muscle strain, tendinopathy, tendinosis, or a muscle- fascial injury. Some embodiments include formulating the platelet composition into a cosmetic composition. Some embodiments include administering the platelet composition to a subject as a cosmetic treatment. In some embodiments, the treatment comprises a dermatological, orthopedic, sports medicine, cosmetic surgery, cosmeceutical, wound healing, dry eye disease, alopecia, ophthalmic surgery, neurosurgery, or general surgery treatment. In some embodiments, the administration comprises an injection. In some embodiments, the administration is to a joint, ankle, elbow, wrist, neck, spine, lower back. In some embodiments, the administration is to an area of poorMintz Docket No.062397-501001WO vascularization in the subject. In some embodiments, the injection is intraarticular, intramuscular (e.g. for hamstring injuries), subdermal injection, or subcutaneous. In some embodiments, the injection comprises microneedling. In some embodiments, the administration is to an eye as an eye drop. In some embodiments, the biofluid samples comprise blood samples, serum samples, plasma samples, or a combination thereof. In some embodiments, the platelet-enriched samples are enriched for platelets by about 1.5x, about 2x, about 2.5x, about 3x, about 3.5x, about 4x, about 4.5x, about 5x, about 5.5x, about 6x, about 6.5x, about 7x, about 7.5x, about 8x, about 8.5x, about 9x, about 9.5x, about 10x, or a range of any two of the aforementioned values, relative to a non-enriched plasma or blood sample. In some embodiments, the platelet-enriched samples are enriched for platelets by about 2-6 times relative to a non-enriched plasma or blood sample. In some embodiments, the platelet composition comprises at 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more platelet-enriched samples. Some embodiments include a platelet composition generated using a method herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0006] FIG. 1 is a flowchart showing an example of donor screening, a first selection,therapeutic biocomponent characterization, and a second selection.

[0007] FIG. 2 is a flowchart showing an example of aspects useful in donor pooling.

[0008] FIG. 3A includes data from a platelet profiling study showing reduction in variability ofcombined cytokine or growth factor measurements in pooled products.

[0009] FIG. 3B includes data from a platelet profiling study showing reduction in variability ofcombined cytokine or growth factor measurements in pooled products.

[0010] FIG. 4A is a plot with data showing an inflammation determination as in platelet-enriched samples from donors of various age groupings.

[0011] FIG. 4B is a plot with data showing an inflammation determination as in platelet-enriched samples from male and female donors grouped by age. DETAILED DESCRIPTION

[0012] Osteoarthritis (OA) is a degenerative joint disease that often involves an entire joint,including cartilage, ligaments, bone, fat, and the synovium. It may lead to pain, stiffness, and loss of mobility, often resulting in joint replacement surgeries. The global burden of OA highlights a significant unmet medical need for effective disease-modifying therapies, as current treatments mainly focus on pain relief with limited impact on disease progression.

[0013] Platelet-rich plasma (PRP) and regenerative medicine may be useful for addressing theunmet need of treatment of subjects with OA. Autologous PRP therapy, which may include aderivative of a patient’s own whole blood containing a high concentration of platelets, has been usedas a regenerative therapy in various medical applications such as osteoarthritis, orthopedics, dental surgery, and wound healing. Platelets are often critical in tissue regeneration due to their ability toMintz Docket No.062397-501001WO release growth factors and cytokines that promote cell migration, proliferation, and angiogenesis. Autologous PRP therapy may provide pain relief in OA patients through anti-inflammatory and tissue regenerative factors such as chemokine (C-C motif) ligand 2 (CCL2), interleukin-1 beta (IL-1β),platelet derived growth factor (PDGF) subunits A, B and C, transforming growth factor β1 (TGF‑β1),insulin-like growth factor 1 (IGF‑1), fibroblast growth factor 2 (FGF‑2), and hepatocyte growth factor(HGF). However, clinical efficacy of autologous PRP is often compromised by variability in platelet quality, leading to inconsistent therapeutic outcomes.

[0014] As such, there are various challenges with autologous PRP. Autologous PRP can be usedin treating osteoarthritis, but its clinical efficacy may be variable due to factors such as donor variability, preparation methods, activation protocols, and administration. These inconsistencies canlimit autologous PRP’s potential as a reliable treatment. Additionally, autologous PRP is expensivefor patients, often not reimbursed by insurance, and may require a complex preparation process that can introduce further variability and quality control issues.

[0015] An improved approach is to use optimized platelet therapy with a focus on characterizingand optimizing various biotherapeutic platelet components as quality control measures to create platelet products such as a shelf-stable, lyophilized platelet drug product for osteoarthritis treatment. Some such methods described herein leverage advanced donor selection, characterization, and platelet pooling techniques to ensure a presence and reduced variability in concentration of some cytokines and growth factors to make certain the production of a more consistent drug substance.

[0016] The disclosure herein provides platelet compositions and methods of making and usingthem. Some methods herein include producing a consistent and optimized platelet-based therapeutic product through the use of donor selection and pooling of platelet concentrates. This disclosure provides a novel method for optimizing platelet-based therapies by addressing the issues of donor variability and therapeutic inconsistency. When implementing specific donor selection criteria and pooling techniques, the methods herein are useful in the production of a more consistent drug substance for treating conditions such as osteoarthritis. Platelet compositions

[0017] Disclosed herein, in some embodiments, are platelet compositions. A platelet compositionmay include a platelet-enriched samples of donor subjects. The platelet composition may be made or generated using a method herein. For example, the platelet composition may be a product of donor selection or platelet sample pooling. The platelet composition may be used in a method such as a method of treatment.

[0018] The platelet composition may include a biofluid sample or combination of biofluidsamples of donor subjects. Biofluid samples may include blood. Blood may be whole blood, or may be fractionated. Biofluid samples may include plasma. Biofluid samples may include serum. In some embodiments, a biofluid sample is a whole blood sample or a plasma sample. A biofluid sample orMintz Docket No.062397-501001WO combination of biofluid samples may have been processed to generate the platelet composition. For example, biofluid samples of donor subjects may have been enriched for platelets, thereby producing platelet-enriched samples. The platelet-enriched samples may be combined in making a platelet composition.

[0019] A platelet composition may include a platelet-enriched sample. In some embodiments, theplatelet composition comprises platelet-enriched samples (e.g. pooled platelet-enriched samples). A platelet composition may include a combination of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 platelet-enriched samples, or a range defined by any 2 of the aforementioned numbers of platelet- enriched samples. In some embodiments, the platelet composition comprises at least 2 platelet- enriched samples. In some embodiments, the platelet composition comprises at least 3 platelet- enriched samples. In some embodiments, the platelet composition comprises at least 4 platelet- enriched samples. In some embodiments, the platelet composition comprises at least 5 platelet- enriched samples. In some embodiments, the platelet composition comprises at least 6 platelet- enriched samples. In some embodiments, the platelet composition comprises at least 7 platelet- enriched samples. In some embodiments, the platelet composition comprises at least 8 platelet- enriched samples. In some embodiments, the platelet composition comprises at least 9 platelet- enriched samples. In some embodiments, the platelet composition comprises at least 10 platelet- enriched samples. In some embodiments, the platelet composition comprises less than 2 platelet- enriched samples. In some embodiments, the platelet composition comprises less than 3 platelet- enriched samples. In some embodiments, the platelet composition comprises less than 4 platelet- enriched samples. In some embodiments, the platelet composition comprises less than 5 platelet- enriched samples. In some embodiments, the platelet composition comprises less than 6 platelet- enriched samples. In some embodiments, the platelet composition comprises less than 7 platelet- enriched samples. In some embodiments, the platelet composition comprises less than 8 platelet- enriched samples. In some embodiments, the platelet composition comprises less than 9 platelet- enriched samples. In some embodiments, the platelet composition comprises less than 10 platelet- enriched samples. Platelet-enriched samples

[0020] A platelet composition may be made from platelet-enriched samples. Some embodimentsrelate to a platelet-enriched sample. A platelet-enriched sample may be generated from a plasma or blood sample, such as a non-enriched plasma or blood sample, from a donor subject. A platelet- enriched sample may be enriched for platelets relative to a non-enriched plasma or blood sample.

[0021] In some embodiments, platelet-enriched samples are enriched for platelets by about 1.5x,about 2x, about 2.5x, about 3x, about 3.5x, about 4x, about 4.5x, about 5x, about 5.5x, about 6x, about 6.5x, about 7x, about 7.5x, about 8x, about 8.5x, about 9x, about 9.5x, about 10x, or a range of any two of the aforementioned values, relative to a non-enriched plasma or blood sample. In someMintz Docket No.062397-501001WO embodiments, a platelet-enriched sample is enriched for platelets by at least 1.5x, at least 2x, at least 2.5x, at least 3x, at least 3.5x, at least 4x, at least 4.5x, at least 5x, at least 5.5x, at least 6x, at least 6.5x, at least 7x, at least 7.5x, at least 8x, at least 8.5x, at least 9x, at least 9.5x, or at least 10x, relative to a non-enriched plasma or blood sample. In some embodiments, a platelet-enriched sample is enriched for platelets by less than 1.5x, less than 2x, less than 2.5x, less than 3x, less than 3.5x, less than 4x, less than 4.5x, less than 5x, less than 5.5x, less than 6x, less than 6.5x, less than 7x, less than 7.5x, less than 8x, less than 8.5x, less than 9x, less than 9.5x, or less than 10x, relative to a non- enriched plasma or blood sample. In some embodiments, the platelet-enriched samples are enriched for platelets by about 2-6 times relative to a non-enriched plasma or blood sample.

[0022] A platelet-enriched sample may be generated through a variety of mechanisms. In someembodiments, the platelet-enriched samples are generated via a series of centrifugation steps or by apheresis of a blood sample. A platelet-enriched sample may be obtained by starting with whole blood which is processed by a series of centrifugations of certain durations and at certain speeds to isolate platelets. A platelet-enriched sample may be obtained by an apheresis machine where a donor is connected to the machine through which their blood flows. The machine may remove platelets from adonor’s body and flow a platelet-reduced blood back into a donor.

[0023] Some embodiments relate to a platelet-rich plasma (PRP) sample. For example, a platelet-enriched sample may be or include a PRP sample. A platelet composition may be or include PRP samples. In some embodiments, the PRP comprises a concentrate of platelets and plasma proteins. In some embodiments, the PRP comprises a concentrate of platelets and plasma proteins. In some embodiments, the PRP comprises a concentrate of platelets and plasma proteins.

[0024] A PRP sample may be or include a concentrate of plasma protein derived from wholeblood, centrifuged to remove red blood cells but retaining platelets. As a concentrated source of plasma, PRP may include multiple growth factors and other cytokines that can stimulate the healing of soft tissues and joints. Various preparation protocols exist, where the underlying principle may be to concentrate platelets to about 2–6 or 3–5 times physiological levels. In some embodiments, a concentrate comprises at least about 2–6 times more platelets than non-enriched plasma. In some embodiments, a concentrate comprises at least about 3–5 times more platelets than non-enriched plasma. Formulations

[0025] Disclosed herein, in some embodiments, are platelet compositions formulated for a usesuch as a pharmaceutical use or cosmetic use. In some embodiments, a platelet composition may be formulated for pharmaceutical use. In some embodiments, a platelet composition may be formulated for cosmetic use.

[0026] Some embodiments include formulating the platelet composition into a pharmaceuticalcomposition. Some embodiments include formulating pooled platelet-enriched samples into aMintz Docket No.062397-501001WO pharmaceutical composition. In some embodiments, the pharmaceutical composition comprises a liquid solution. In some embodiments, the pharmaceutical composition comprises a lyophilized platelet composition.

[0027] Some embodiments include formulating the platelet composition into a cosmeticcomposition. Some embodiments include formulating pooled platelet-enriched samples into a cosmetic composition. In some embodiments, the cosmetic composition comprises a liquid solution. In some embodiments, the cosmetic composition comprises a lyophilized platelet composition.

[0028] A platelet composition may include an additive or additives. An additive may include asalt. An additive may include a buffer. An additive may include a sugar such as a monosaccharide, disaccharide, or polysaccharide. Multiple additives may be included. For example, a platelet composition may include a buffer and an added sugar.

[0029] A platelet composition may be in liquid form. For example, the platelet composition mayinclude an aqueous carrier. An aqueous carrier may include water or another solvent. The platelet composition may include water. A platelet composition may include a buffer. The buffer may be in solution. The buffer may be dissolved in water.

[0030] A platelet composition may be lyophilized. The lyophilized platelet composition mayinclude an additive. The additive may be lyophilized with platelets of the platelet composition. The lyophilized platelet composition may include a buffer that is lyophilized with platelets. The lyophilized platelet composition may be reconstituted in a solvent. In some embodiments, a lyophilized platelet composition is formulated for reconstitution with a reconstitution buffer by a medical practitioner.

[0031] In some embodiments, a platelet composition is formulated for injection. In someembodiments, pooled platelet-enriched samples are formulated for injection. Some embodiments relate to a needle comprising a platelet composition, ready for injection.

[0032] In some embodiments, a platelet composition is formulated for intravenousadministration. In some embodiments, pooled platelet-enriched samples are formulated for intravenous administration. Some embodiments relate to an IV comprising a platelet composition. Making platelet compositions

[0033] Disclosed herein, in some embodiments, are methods of making a platelet composition.Some embodiments relate to a method for producing a platelet-based therapeutic product, comprising: selecting platelet donors based on predefined cytokine and growth factor profiles, age, disease status, and a systemic inflammation measurement such as plasma C-reactive protein concentration (CRP). Some embodiments involve donor selection or making a donor selection decision. A method of making a platelet composition may include donor selection. Some embodiments involve pooling of samples such as platelet-enriched samples of selected donors. A method of making a platelet composition may include pooling the platelet-enriched samples of selected donors.Mintz Docket No.062397-501001WO

[0034] Provided herein, in some embodiments, are methods of making a platelet composition,comprising: determining characteristics of each donor subject of a group of donor subjects, the characteristics comprising: measurements of a systemic inflammation marker such as CRP and one or more cytokines or growth factors in a biofluid sample of the donor subject, and an age and disease status of the donor subject; and based on the characteristics, pooling platelet-rich plasma (PRP, an example of a platelet enriched sample) samples of the donor subjects to obtain pooled PRP samples.

[0035] Some embodiments include, for a donor of a group of donors: determining an age of thedonor from the donor information, determining a disease status of the donor from the donor information or from disease status measurements obtained from one or more of the biofluid samples, obtaining a systemic inflammation measurement for the donor from the donor information or from a systemic inflammation measurement obtained from one or more of the biofluid samples, obtaining cytokine or growth factor measurements for the donor from a platelet-enriched sample of the donor, pooling the platelet-enriched sample of the donor with other platelet-enriched samples when the donor age is within a predetermined age range, the donor disease status is free of a predetermined selection of diseases, the systemic inflammation measurement is below a predetermined systemic inflammation threshold, and the cytokine or growth factor measurements are within predetermined cytokine or growth factor ranges, thereby generating a platelet composition comprising pooled platelet-enriched samples of the group of human donors, and not pooling the platelet-enriched sample of the donor with the other platelet-enriched samples when the donor age is not within the predetermined age range, the donor disease status is not free of the predetermined selection of diseases, the systemic inflammation measurement is not below the predetermined systemic inflammation threshold, or the cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges.

[0036] Provided herein, in some embodiments, are methods of making a platelet composition.Some embodiments of the method include: obtaining donor information for a group of human donors; optionally obtaining biofluid samples of the group of human donors; obtaining platelet-enriched samples of the group of human donors; and for a donor of the group of donors: determining an age of the donor from the donor information, determining a disease status of the donor from the donor information or from disease status measurements obtained from one or more of the biofluid samples, obtaining a systemic inflammation measurement for the donor from the donor information or from a systemic inflammation measurement obtained from one or more of the biofluid samples, obtaining cytokine or growth factor measurements for the donor from a platelet-enriched sample of the donor, pooling the platelet-enriched sample of the donor with other platelet-enriched samples when the donor age is within a predetermined age range, the donor disease status is free of a predetermined selection of diseases, the systemic inflammation measurement is below a predetermined systemic inflammation threshold, and the cytokine or growth factor measurements are within predetermined cytokine or growth factor ranges, thereby generating a platelet composition comprising pooled platelet-enriched samples of the group of human donors, and not pooling the platelet-enriched sample of the donor withMintz Docket No.062397-501001WO the other platelet-enriched samples when the donor age is not within the predetermined age range, the donor disease status is not free of the predetermined selection of diseases, the systemic inflammation measurement is not below the predetermined systemic inflammation threshold, or the cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges.

[0037] Provided herein, in some embodiments, are methods of making a platelet composition,comprising: obtaining donor samples or donor information for a group donors; and for each donor of the group of donors: determining a donor age from the donor information, determining a donor disease status from the donor information, obtaining a donor systemic inflammation measurement (e.g. a donor CRP measurement) from one or more donor samples or from the donor information, and obtaining a cytokine or growth factor measurement from the one or more donor samples or from the donor information, pooling a platelet-enriched plasma sample from the donor with other platelet- enriched plasma samples when the donor age is within a predetermined age range, the donor disease status is free of a predetermined selection of diseases, the systemic inflammation measurement is within a predetermined systemic inflammation measurement range (e.g. a predetermined CRP measurement range), and the cytokine or growth factor measurement is within a predetermined cytokine or growth factor range, and not pooling the platelet-enriched plasma sample from the donor with the other platelet-enriched plasma samples when the donor age is not within the predetermined age range, the donor disease status is not free of the predetermined selection of diseases, the systemic inflammation measurement is not within the predetermined systemic inflammation range, or the cytokine or growth factor measurement is not within the predetermined cytokine or growth factor range. The disease status may be further obtained from an assay performed on a donor sample.

[0038] Some embodiments include further processing of pooled samples. Some embodimentsinclude enriching pooled platelet-enriched samples further. Enrichment may be performed to increase a platelet concentration of a platelet composition. The further enrichment may include centrifugation. The further enrichment may include removal of cells such as erythrocytes or leukocytes. Some embodiments include enriching the platelet composition further by centrifuging the pooled platelet- enriched samples and removing erythrocytes and leukocytes.

[0039] Further processing of pooled samples may include combining pooled platelet-enrichedsamples with a buffer solution. Further processing of pooled samples may include combining pooled platelet-enriched samples with a buffer solution. Some embodiments include combining a platelet composition with a buffer.

[0040] Some embodiments include combining pooled platelet-enriched samples with amonosaccharide, disaccharide, or polysaccharide. Some embodiments include combining a platelet composition with a monosaccharide, disaccharide, or polysaccharide.

[0041] The pooled samples may be reconstituted. Further processing of pooled samples mayinclude lyophilizing the pooled samples. Pooled samples may be formulated into a platelet composition herein. Pooled samples may be packaged. A platelet composition may be packaged.Mintz Docket No.062397-501001WO Lyophilization

[0042] Disclosed herein, in some embodiments, are methods that include a dehydration processsuch as lyophilization. A platelet composition may be lyophilized. Some embodiments include lyophilizing pooled platelet-enriched samples. Some embodiments include lyophilizing a platelet composition. In some embodiments, a pooled product is used as a drug substance to be further processed into a lyophilized drug product. In some embodiments, a pooled product is used as a drug substance to be further processed into a lyophilized drug product for treating osteoarthritis by intraarticular injection. Lyophilization may include freezing and lowering of pressure, thereby removing ice by sublimation.

[0043] Some embodiments include obtaining cytokine or growth factor measurements (e.g.second cytokine or growth factor measurements) from a platelet composition before the lyophilization. Some embodiments include lyophilizing a platelet composition when cytokine or growth factor measurements are within predetermined cytokine or growth factor ranges. Some embodiments include not lyophilizing a platelet composition when cytokine or growth factor measurements are not within predetermined cytokine or growth factor ranges. Some embodiments include lyophilizing the platelet composition when the second cytokine or growth factor measurements are within the predetermined cytokine or growth factor ranges, and not lyophilizing the platelet composition when the second cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges.

[0044] Some embodiments include lyophilizing a platelet composition when combined cytokineor growth factor measurements (e.g. a Sentinel index) are within predetermined a combined cytokine or growth factor range. Some embodiments include not lyophilizing a platelet composition when combined cytokine or growth factor measurements are not within a predetermined combined cytokine or growth factor range. Some embodiments include lyophilizing a platelet composition when a second combined cytokine or growth factor measurement is within a predetermined combined cytokine or growth factor range, and not lyophilizing the platelet composition when the second combined cytokine or growth factor measurement is not within the combined predetermined cytokine or growth factor range.

[0045] Some embodiments include reconstituting lyophilized pooled platelet-enriched samples.Some embodiments include reconstituting the lyophilized platelet composition. Reconstitution may involve adding a wetting reagent such as water or a buffer. Reconstitution may include mixing. Packaging

[0046] Some embodiments include packaging a platelet composition. A packaged plateletcomposition may be included in a kit. The packaged platelet composition may be in liquid form. The packaged platelet composition may be lyophilized.Mintz Docket No.062397-501001WO

[0047] Some embodiments include obtaining second or third cytokine or growth factormeasurements from the platelet composition before the packaging. Some embodiments include packaging the platelet composition when the second or third cytokine or growth factor measurements are within the predetermined cytokine or growth factor ranges, and not packaging the platelet composition when the second or third cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges. Donor selection

[0048] Disclosed herein, in some embodiments, are methods that include donor selection. Donorselection may include selecting a donor. Donor selection may include selecting donors. Donor selection may be included in a method such as a method of making a platelet composition. Some embodiments of donor selection relate to selecting a possible donor to be a donor. Selecting a possible donor to be a donor may include or be followed by obtaining a biofluid sample such as a blood sample from the donor. The blood sample may be processed to obtain a platelet-enriched sample. Some embodiments of donor selection relate to selecting a donor (e.g. from a group of donors). Selecting a donor may include or be followed by obtaining a platelet-enriched sample of the selected donor, obtaining a measurement or measurements such as a cytokine or growth factor measurement or cytokine or growth factor measurements from a platelet-enriched sample of the donor, pooling of theselected donor’s platelet-enriched sample with other platelet-enriched samples of donors, orgenerating a platelet composition from a platelet-enriched sample of the selected donor.

[0049] In some embodiments, a method begins with identifying and selecting platelet donorsbased on specific cytokine and growth factors that have tissue regenerative therapeutic utility. This is useful for producing a consistent drug substance. Donors may be screened using an algorithm that evaluates their platelet cytokine levels, including IGF-1, CCL-2, TNF, IL-1B, PDGF-BB, EGF,CTGF, FGF-2, HGF, TSP-1, TGF-β1, and, PF4, a selection thereof, or other growth factors orcytokines herein that influence the therapeutic efficacy of the final product. In some embodiments, only donors whose profiles meet predefined ranges of selected cytokines and growth factors are selected for further processing. In addition to cytokine levels that represent regenerative potential of platelets, additional criteria such as age and inflammatory status of the donor may be used to optimize donor selection, for example in a case where the growth factors are independent of age and inflammatory markers.

[0050] In addition to cytokine levels, the age of the donor may be useful in a selection process.Platelet function may change significantly with age. For instance, aging may be associated with increased platelet aggregability, a reduced platelet count, and a prothrombotic state in older individuals, particularly those over the age of 60. Platelets from older donors may tend to be more prone to activation and less sensitive to inhibition, which can negatively impact the consistency and efficacy of platelet-based therapies.Mintz Docket No.062397-501001WO

[0051] Furthermore, platelet turnover rates and oxidative stress may be elevated in olderindividuals, further contributing to platelet hyperactivity and the likelihood of undesirable pro- inflammatory factors. Therefore, to optimize the therapeutic potential of the platelet product and ensure consistency in key biological factors, donors above a certain age threshold (e.g.60 or 40 years of age) may be excluded from a donor pool. In some embodiments, donors are of an age less than 60. In some embodiments, donors are of an age less than 40 years of age.

[0052] By incorporating age as a selection criterion, variability in platelet function and cytokineprofiles may be reduced. This is useful in producing a more reliable and effective therapeutic product for conditions such as osteoarthritis.

[0053] In addition to cytokine profiles and age, a critical selection criterion may be implementedbased on a systemic inflammatory marker such as a C-reactive protein (CRP) measurement. The systemic inflammation measurement (e.g. CRP) measurement may be obtained in a plasma sample. Elevated CRP levels may correspond with chronic inflammation or a disease state such as an inflammatory diseases, cardiovascular disease, or type 2 diabetes mellitus. The presence of such inflammation or an inflammatory disease state may compromise the quality of platelet concentrates. CRP or another systemic inflammation marker may be useful as an acute-phase reactant whose elevated levels reflect underlying inflammation, which may negatively impact the regenerative properties of platelet-derived growth factors and cytokines essential for therapeutic efficacy. To minimize variability and ensure consistency in the therapeutic product, donors with systemic inflammation measurements equal to or greater than a threshold (e.g. CRP concentrations equal to or greater than a threshold such as 3.0 mg / L) may be excluded. Implementing the systemic inflammation threshold (e.g. CRP threshold) is useful for further reducing donor-related variability and enhancing the consistency and quality of a platelet-based therapeutic product.

[0054] FIG. 1 includes example donor screening processes, including: health and safetyscreening, therapeutic biocomponent (TB) characterization, and a second selection based on the TB characterization. Any one or a combination of these aspects may be used in donor selection. For example, a donor selection may involve health and safety screening. A donor selection may involve TB characterization. A donor selection may involve selection based on TB characterization. A donor selection may involve health and safety screening, TB characterization, and selection based on the TB characterization.

[0055] In some embodiments, patient selection involves obtaining a biofluid sample. Thebiofluid sample may be obtained from a donor or potential donor. Some embodiments include obtaining multiple biofluid samples. In some embodiments, the biofluid samples comprise blood samples, serum samples, plasma samples, or a combination thereof.

[0056] In some embodiments, obtaining a platelet-enriched sample of the donor is optional. Forexample, some embodiments do not include obtaining a platelet-enriched sample of a possible donor. In some embodiments, a platelet-enriched sample of a donor or potential donor is obtained when theMintz Docket No.062397-501001WO donor or potential donor age is within the predetermined age range. In some embodiments, a platelet-enriched sample of a donor or potential donor is obtained when the donor or potential donor’s diseasestatus is free of the predetermined selection of diseases. In some embodiments, a platelet-enriched sample of a donor or potential donor is obtained when a systemic inflammation measurement of the donor or potential donor is below a predetermined systemic inflammation threshold. In some embodiments, a platelet-enriched sample of a donor or potential donor is not obtained when the donor or potential donor age is not within the predetermined age range. In some embodiments, a platelet-enriched sample of a donor or potential donor is not obtained when the donor or potential donor’sdisease status is not free the predetermined selection of diseases. In some embodiments, a platelet- enriched sample of a donor or potential donor is not obtained when the systemic inflammation measurement is not below the predetermined systemic inflammation threshold. In some embodiments, when a platelet-enriched sample of a donor or potential donor is not obtained, no platelet-enriched sample of the donor is pooled with other platelet-enriched samples. In some embodiments, obtaining the platelet-enriched sample of the donor is optional, and wherein: the platelet-enriched sample of the donor is obtained when the donor age is within the predetermined age range, the donor disease status is free of the predetermined selection of diseases, and the systemic inflammation measurement is below the predetermined systemic inflammation threshold; and the platelet-enriched sample of the donor is not obtained when the donor age is not within the predetermined age range, the donor disease status is not free the predetermined selection of diseases, or the systemic inflammation measurement is not below the predetermined systemic inflammation threshold, wherein when the platelet-enriched sample of the donor is not obtained, no platelet-enriched sample of the donor is pooled with the other platelet-enriched samples.

[0057] In some embodiments, a donor is a vertebrate. In some embodiments, the donor subjectsare vertebrate donor subjects. In some embodiments, a donor is an animal. In some embodiments, the donor subjects are animal donor subjects. In some embodiments, a donor is a mammal. In some embodiments, the donor subjects are mammalian donor subjects. In some embodiments, a donor is a primate. In some embodiments, the donor subjects are primate donor subjects. In some embodiments, a donor is a human. In some embodiments, the donor subjects are human donor subjects.

[0058] In some embodiments, a donor is male. In some embodiments, a group of donors includeor consist of male donors. In some embodiments, a donor is female. In some embodiments, a group of donors include or consist of female donors. In some embodiments, a group of donors includes a combination of male and female donors.

[0059] Some embodiments include selecting a donor from a candidate donor or group ofcandidate donors. The selection may be based on a factor or combination of factors such as age, disease status, or a systemic inflammation measurement. Selecting a donor may be based on acandidate donor’s age. Selecting a donor may be based on a candidate donor’s disease status.Selecting a donor may be based on a candidate donor’s systemic inflammation measurement.Mintz Docket No.062397-501001WOSelecting a donor may be based on a candidate donor’s age, disease status, and systemic inflammationmeasurement. A selected donor may be allowed to donate a biofluid such as a blood or plasma sample. Some embodiments include collecting a biofluid from a selected donor. The biofluid may be processed into a platelet-enriched sample, or may be further processed into a platelet composition.

[0060] A method (e.g. a donor selection method or a method of making a platelet compositionthat includes donor selection) may include obtaining information of a donor or potential donor. A method may include obtaining information of a group of donor or group of potential donor. The information may be obtained directly, for example from a question or survey. The information may be obtained indirectly, for example by receiving a report.

[0061] A method may include obtaining information of a potential donor or group of potentialdonors. Information may include an age. Information may include a disease status. Information may include a systemic inflammation measurement. Information may include a cytokine or growth factor measurement. Some embodiments include obtaining information for a group of potential human donors. A method may include obtaining biofluid samples from potential donors. Some embodiments include obtaining biofluid samples of a group of potential human donors. A method may include obtaining platelet-enriched samples from potential donors. Some embodiments include obtaining platelet-enriched samples of a group of potential human donors. Some embodiments include obtaining a PRP sample from a subject. Some embodiments include obtaining PRP samples from subjects.

[0062] The method may include using potential donor information or biofluid samples from thedonors to make a potential donor selection decision. The potential donor selection decision may include selecting the potential donor to be a donor. If a potential donor is selected to be a donor, a biofluid sample may be obtained from the selected potential donor. Selecting a potential donor may include selecting a platelet-enriched sample of the potential donor for further analysis, such as further analysis in making a further donor selection decision. The potential donor selection decision may include rejecting the potential donor. Rejecting a potential donor may include not allowing the potential donor to donate a biofluid sample. Rejecting a potential donor may include not obtaining a biofluid sample from the potential donor.

[0063] A method may include obtaining donor information and biofluid samples from donors. Amethod may include obtaining donor information from donors. Some embodiments include obtaining donor information for a group of human donors. Donor information may include age, disease status, a systemic inflammation measurement, or a combination thereof. Donor information may include a cytokine or growth factor measurement. A method may include obtaining biofluid samples from donors. Some embodiments include obtaining biofluid samples of a group of human donors. A method may include obtaining platelet-enriched samples from donors. Some embodiments include obtaining platelet-enriched samples of a group of human donors.

[0064] The method may include using the donor information or biofluid samples from the donorsto make a donor selection decision. The donor selection decision may include selecting the donor.Mintz Docket No.062397-501001WO Selecting a donor may include selecting a platelet-enriched sample of the donor for further analysis, such as further analysis in making a further donor selection decision. Selecting a donor may include selecting a platelet-enriched sample of the donor for pooling with other platelet-enriched samples. The donor selection decision may include rejecting the donor. Rejecting a donor may include rejecting a platelet-enriched sample of the donor for further analysis, such as further analysis in donor selection decision. Rejecting a donor may include rejecting a platelet-enriched sample of the donor for pooling with other platelet-enriched samples.

[0065] Some embodiments include determining an age of a donor from the donor information.Some embodiments include determining a disease status of the donor from the donor information or from disease status measurements obtained from one or more of the biofluid samples. Some embodiments include determining a disease status of the donor from the donor information. Some embodiments include determining a disease status of the donor from disease status measurements obtained from one or more of the biofluid samples. Some embodiments include obtaining a systemic inflammation measurement for the donor from the donor information or from a systemic inflammation measurement obtained from one or more of the biofluid samples. Some embodiments include obtaining a systemic inflammation measurement for the donor from the donor information. Some embodiments include obtaining a systemic inflammation measurement for the donor from a systemic inflammation measurement obtained from one or more of the biofluid samples.

[0066] Some embodiments include obtaining cytokine or growth factor measurements. Thecytokine or growth factor measurements may include a cytokine measurement. The cytokine or growth factor measurements may include cytokine measurements. The cytokine or growth factor measurements may include a growth factor measurement. The cytokine or growth factor measurements may include growth factor measurements. Some embodiments include obtaining a cytokine measurement. Some embodiments include obtaining cytokine measurements. Some embodiments include obtaining a growth factor measurement. Some embodiments include obtaining growth factor measurements. Some embodiments include obtaining cytokine or growth factor measurements for the donor from a platelet-enriched sample of the donor. A measurement may include a readout or amount. Donor age

[0067] Donor selection may be based on age (e.g. based at least in part on age). A method ofmaking a platelet composition may include donor selection based on age. Some embodiments involve pooling of samples such as platelet-enriched samples of selected donors within a predetermined age range. A method of making a platelet composition may include pooling the platelet-enriched samples of selected donors where the selected donors have an age within the predetermined age range.

[0068] In some embodiments, the predetermined age range includes an age of at least 10 years, atleast 12 years, at least 14 years, at least 16 years, at least 18 years, at least 20 years, at least 22 years,Mintz Docket No.062397-501001WO at least 24 years, at least 26 years, at least 28 years, or at least 30 years. In some embodiments, the predetermined age range includes an age below 60 years, below 58 years, below 56 years, below 54 years, below 52 years, below 50 years, below 48 years, below 46 years, below 44 years, below 42 years, below 40 years, below 38 years, below 36 years, below 34 years, or below 32 years, or below 30 years.

[0069] Some embodiments include selecting a subject within a predetermined age. Someembodiments include rejecting a subject outside a predetermined age.

[0070] In some embodiments, pooled platelet-enriched samples are of donor subjects of an agerange. In some embodiments, platelet-enriched samples of donor subjects outside the age range are excluded from the pooled platelet-enriched samples. In some embodiments, the pooled platelet- enriched samples are of donor subjects of an age range, and platelet-enriched samples of donor subjects outside the age range are excluded from the pooled platelet-enriched samples. In some embodiments, pooled PRP samples are of donor subjects of an age range. In some embodiments, PRP samples of donor subjects outside the age range are excluded from the pooled PRP samples. In some embodiments, the pooled PRP samples are of donor subjects of an age range, and PRP samples of donor subjects outside the age range are excluded from the pooled PRP samples. Donor disease status

[0071] Donor selection may be based on disease status (e.g. based at least in part on diseasestatus). A method of making a platelet composition may include donor selection based on disease status. Some embodiments involve pooling of samples such as platelet-enriched samples of selected donors of a predetermined disease status. A method of making a platelet composition may include pooling the platelet-enriched samples of selected donors where the selected donors are of a predetermined disease status.

[0072] A disease status may include the presence or absence of a predetermined disease status. Adisease status may include a presence of any one of a predetermined selection of diseases. A disease status may include an absence of a predetermined selection of diseases. A predetermined disease status may include a lack of a predetermined selection of diseases.

[0073] A predetermined selection of diseases may include an infectious disease, cancer,autoimmune disease, cardiovascular disease, or a combination thereof. The predetermined selection of diseases may include cancer. The predetermined selection of diseases may include an autoimmune disease. The predetermined selection of diseases may include a cardiovascular disease. The predetermined selection of diseases may include an infectious disease. In some embodiments, a predetermined selection of diseases comprises infectious disease, cancer, autoimmune disease, and cardiovascular disease.

[0074] Examples of infectious diseases may include human immunodeficiency virus, human T-cell lymphotropic virus, hepatitis B, hepatitis C, syphilis, West Nile virus, or Chagas disease. TheMintz Docket No.062397-501001WO predetermined selection of diseases may include human immunodeficiency virus. The predetermined selection of diseases may include human T-cell lymphotropic virus. The predetermined selection of diseases may include hepatitis B. The predetermined selection of diseases may include hepatitis C. The predetermined selection of diseases may include syphilis. The predetermined selection of diseases may include West Nile virus. The predetermined selection of diseases may include Chagas disease. In some embodiments, the predetermined selection of diseases comprises human immunodeficiency virus, human T-cell lymphotropic virus, hepatitis B, hepatitis C, syphilis, West Nile virus, and Chagas disease.

[0075] At least some of the predetermined selection of diseases may be identified from donorinformation or from potential donor information. An infectious disease, cancer, autoimmune disease, cardiovascular disease, or a combination thereof may be identified from donor information or from potential donor information. Cancer may be identified from donor information or from potential donor information. An autoimmune disease may be identified from donor information or from potential donor information. A cardiovascular disease may be identified from donor information or from potential donor information. An infectious disease may be identified from donor information or from potential donor information. An infectious disease, cancer, autoimmune disease, and cardiovascular disease may be identified from donor information or from potential donor information. At least some of the predetermined selection of diseases may be identified from disease status measurements. At least some of the predetermined selection of diseases may be identified from disease status measurements obtained from one or more of the biofluid samples.

[0076] Human immunodeficiency virus, human T-cell lymphotropic virus, hepatitis B, hepatitisC, syphilis, West Nile virus, or Chagas disease may be identified from donor information or from potential donor information. Human immunodeficiency virus, human T-cell lymphotropic virus, hepatitis B, hepatitis C, syphilis, West Nile virus, or Chagas disease may be identified from disease status measurements. Disease status measurements may be obtained from one or more of the biofluid samples. Systemic inflammation

[0077] Donor selection may be based on systemic inflammation (e.g. based at least in part onsystemic inflammation). A method of making a platelet composition may include donor selection based on systemic inflammation. Some embodiments involve pooling of samples such as platelet- enriched samples of selected donors lacking systemic inflammation. A method of making a platelet composition may include pooling the platelet-enriched samples of selected donors where the selected donors lack systemic inflammation.

[0078] Systemic inflammation may be determined through a systemic inflammationmeasurement. A systemic inflammation measurement may include a readout or amount of a systemic inflammation marker. A systemic inflammation marker may include C-reactive protein (CRP) or anMintz Docket No.062397-501001WO erythrocyte sedimentation rate (ESR). A systemic inflammation marker may include CRP. A systemic inflammation marker may include an ESR. Some examples of systemic inflammation markers may include a tumor necrosis factor (TNF), liver enzymes, amyloid, Tau, neurofilament light polypeptide (Nfl), or glial fibrillary acidic protein (GFAP), any of which may be useful to include in a systemic inflammation measurement.

[0079] In some embodiments, a systemic inflammation measurement comprises a CRPmeasurement. In some embodiments, a predetermined systemic inflammation threshold comprises a predetermined CRP threshold. In some embodiments, the systemic inflammation measurement comprises a CRP measurement, and the predetermined systemic inflammation threshold comprises a predetermined CRP threshold. A CRP measurement may be obtained in a blood sample. A CRP measurement may be obtained in a biofluid sample. A CRP measurement may be obtained in a plasma sample.

[0080] In some embodiments, a predetermined CRP threshold is 0 mg / L (e.g. undetectable), 0.5mg / L, 1 mg / L, 1.5 mg / L, 2 mg / L, 2.5 mg / L, 3 mg / L, 3.5 mg / L, 4 mg / L, 4.5 mg / L, 5 mg / L, 5.5 mg / L, 6 mg / L, 6.5 mg / L, 7 mg / L, 7.5 mg / L, 8 mg / L, 8.5 mg / L, 9 mg / L, 9.5 mg / L, or 10 mg / L. In some embodiments, a predetermined CRP threshold is about 0 mg / L, about 0.5 mg / L, about 1 mg / L, about 1.5 mg / L, about 2 mg / L, about 2.5 mg / L, about 3 mg / L, about 3.5 mg / L, about 4 mg / L, about 4.5 mg / L, about 5 mg / L, about 5.5 mg / L, about 6 mg / L, about 6.5 mg / L, about 7 mg / L, about 7.5 mg / L, about 8 mg / L, about 8.5 mg / L, about 9 mg / L, about 9.5 mg / L, or about 10 mg / L. In some embodiments, the predetermined CRP threshold is any detectable amount. In some embodiments, the predetermined CRP threshold is at least 0 mg / L, at least 0.5 mg / L, at least 1 mg / L, at least 1.5 mg / L, at least 2 mg / L, at least 2.5 mg / L, at least 3 mg / L, at least 3.5 mg / L, at least 4 mg / L, at least 4.5 mg / L, at least 5 mg / L, at least 5.5 mg / L, at least 6 mg / L, at least 6.5 mg / L, at least 7 mg / L, at least 7.5 mg / L, at least 8 mg / L, at least 8.5 mg / L, at least 9 mg / L, at least 9.5 mg / L, or at least 10 mg / L. In some embodiments, the predetermined CRP threshold is less than 0.5 mg / L, less than 1 mg / L, less than 1.5 mg / L, less than 2 mg / L, less than 2.5 mg / L, less than 3 mg / L, less than 3.5 mg / L, less than 4 mg / L, less than 4.5 mg / L, less than 5 mg / L, less than 5.5 mg / L, less than 6 mg / L, less than 6.5 mg / L, less than 7 mg / L, less than 7.5 mg / L, less than 8 mg / L, less than 8.5 mg / L, less than 9 mg / L, less than 9.5 mg / L, or less than 10 mg / L. Such CRP thresholds may relate to a CRP measurement in plasma.

[0081] In some embodiments, a systemic inflammation measurement comprises an ESRmeasurement. In some embodiments, a predetermined systemic inflammation threshold comprises a predetermined ESR threshold. In some embodiments, the systemic inflammation measurement comprises an ESR measurement, and the predetermined systemic inflammation threshold comprises a predetermined ESR threshold.

[0082] ESR may be determined by the following: a technician places red blood cells into a tall,thin tube and check how far they fall in 1 hour, where inflammation results in abnormal proteins in the blood that form the red blood cells into clumps that move faster relative to single blood cells. Thus, aMintz Docket No.062397-501001WO higher ESR may correspond with increased systemic inflammation. Normal ESR ranges may be as follows, which may be useful in determining a systemic inflammation threshold for donor selection: 0 to 15 mm / hour in men younger than 50 0 to 20 mm / hour in men older than 50 0 to 20 mm / hour in women younger than 50 0 to 30 mm / hour for women older than 50 0 to 10 mm / hour for a child

[0083] In some embodiments, a predetermined ESR threshold is 10 mm / hour, 15 mm / hour, 20mm / hour, 25 mm / hour, 30 mm / hour, 35 mm / hour, or 40 mm / hour. In some embodiments, a predetermined ESR threshold is about 10 mm / hour, about 15 mm / hour, about 20 mm / hour, about 25 mm / hour, about 30 mm / hour, about 35 mm / hour, or about 40 mm / hour. In some embodiments, a predetermined ESR threshold is at least 10 mm / hour, at least 15 mm / hour, at least 20 mm / hour, at least 25 mm / hour, at least 30 mm / hour, at least 35 mm / hour, or at least 40 mm / hour. In some embodiments, a predetermined ESR threshold is less than 10 mm / hour, less than 15 mm / hour, less than 20 mm / hour, less than 25 mm / hour, less than 30 mm / hour, less than 35 mm / hour, or less than 40 mm / hour. Sentinel index

[0084] Donor selection may be based on a cytokine or growth factor measurement (e.g. based atleast in part on a cytokine or growth factor measurement). A cytokine or growth factor measurement may include cytokine or growth factor measurements. A method of making a platelet composition may include donor selection based on a cytokine or growth factor measurement. Some embodiments involve pooling of samples such as platelet-enriched samples of selected donors having cytokine or growth factor measurements within predetermined cytokine or growth factor ranges. A method of making a platelet composition may include pooling the platelet-enriched samples of selected donors where the selected donors are of selected donors having cytokine or growth factor measurements within predetermined cytokine or growth factor ranges.

[0085] Any one of several platelet constituents may have beneficial effects. For example, severalplatelet constituents have been implicated in beneficial effects of autologous platelet-rich plasma therapy for osteoarthritis. Some examples are provided in Andia, Nat. Rev. Rheumatol 2013, which is incorporated herein by reference for its description of chemokines / cytokines, small molecules, growth factors, adhesive proteins, proteases / antiproteases, and in its entirety. Some such chemokines / cytokines, small molecules, growth factors, adhesive proteins, or proteases / antiproteases may be useful in a platelet composition, or may be useful as determinants in cytokine or growth factor measurements herein.Mintz Docket No.062397-501001WO

[0086] Platelet proteins likely associated with autologous PRP’s therapeutic benefits areprovided herein. Some beneficial proteins, referred to here as Sentinels, have been selected for use here based on their roles in physiological processes such as the repair and / or regeneration of cartilage, bone, vasculature, and the synovium as well as their ability to reduce inflammation and beneficially regulate the immune system. Some examples of such Sentinel proteins are provided in Table 1. At least some of these proteins, when present in a platelet-enriched sample such as platelet-rich plasma (PRP), may contribute to the therapeutic, regenerative, reparative, and anti-inflammatory effects in PRP treatments, such as for knee osteoarthritis. Their actions may alleviate symptoms, reduce inflammation, and promote repair of damaged tissues, for example in osteoarthritic joints. The Sentinel proteins are examples of cytokines or growth factors.

[0087] While other therapeutic factors may also be provided by platelets, an allogenic plateletformulation lacking the 6 Sentinel proteins identified herein or having concentrations significantly below normal physiologic levels is less likely to be useful for therapeutic use. A presence of Sentinel proteins at or above healthy concentrations is more likely to be beneficial. Therefore, such Sentinel proteins may be useful in donor selection.

[0088] A presence of a useful amount of a Sentinel protein may be beneficial in a disease ordisorder such as osteoarthritis (OA, e.g. knee OA). A presence of a useful amount of a Sentinel protein may provide a benefit through promoting wound healing, tissue regeneration, tissue repair (e.g. synovial tissue repair), cartilage repair, cartilage regeneration, regeneration of other joint tissues, reducing pain, improving joint function (e.g. knee function), reducing tissue or joint inflammation (e.g. knee inflammation), or modulation of an immune response. A presence of a useful amount of a Sentinel protein may provide a benefit through stimulating cell proliferation, cell motility (e.g. chemotaxis), cell differentiation, morphogenesis, angiogenesis, synthesis of extracellular matrix components such as collagen or proteoglycans, activation of stem cells, or stem cell differentiation into chondrocytes or synoviocytes. A presence of a useful amount of a Sentinel protein may provide a benefit by inhibiting catabolic processes that lead to cartilage degradation, or providing anti- inflammatory effects.

[0089] Disclosed herein, in some embodiments, are cytokine or growth factor measurements. Insome embodiments, the cytokine or growth factor measurements comprise a chemokine C-C motif ligand 2 (CCL-2) measurement. In some embodiments, the cytokine or growth factor measurements comprise a connective tissue growth factor (CTGF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a fibroblast growth factor (FGF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a fibroblast growth factor 2 (FGF-2) measurement. In some embodiments, the cytokine or growth factor measurements comprise a hepatocyte growth factor (HGF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a histamine measurement. In some embodiments, the cytokine or growth factor measurements comprise an insulin-like growth factor 1 (IGF-1) measurement. In someMintz Docket No.062397-501001WO embodiments, the cytokine or growth factor measurements comprise an interleukin-1 beta (IL-1β) measurement. In some embodiments, the cytokine or growth factor measurements comprise a platelet- derived growth factor (PDGF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a platelet-derived growth factor BB (PDGF-BB) measurement. In some embodiments, the cytokine or growth factor measurements comprise a platelet factor 4 (PF4) measurement. In some embodiments, the cytokine or growth factor measurements comprise a prostaglandin E2 measurement. In some embodiments, the cytokine or growth factor measurementscomprise a transforming growth factor-beta (TGF-β) measurement. In some embodiments, thecytokine or growth factor measurements comprise a transforming growth factor-beta 1 (TGF-β1) measurement. In some embodiments, the cytokine or growth factor measurements comprise a tumor necrosis factor (TNF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a thrombospondin-1 (TSP-1) measurement. In some embodiments, the cytokine or growth factor measurements comprise an epidermal growth factor (EGF) measurement. In some embodiments, the cytokine or growth factor measurements comprise a vascular endothelial growth factor (VEGF) measurement. Any cytokine or growth factor in FIG.1 (referred to as TBs in the figure), or combination of cytokines or growth factors in FIG.1, may be measured to obtain cytokine or growth factor measurements. Any cytokine or growth factor in Table 1, or combination of cytokines or growth factors in Table 1, may be measured to obtain cytokine or growth factor measurements. Such cytokines and growth factors may be measured and used in donor selection, or in platelet composition monitoring.

[0090] The cytokine or growth factor measurements may include measurements of 1, 2, 3, 4, 5,6, 7, 8, 9, or 10 cytokines or growth factors, or a range thereof. The cytokine or growth factor measurements may include measurements of at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 cytokines or growth factors. The cytokine or growth factor measurements may include measurements of less than 2, less than 3, less than 4, less than 5, less than 6, less than 7, less than 8, less than 9, or less than 10 cytokines or growth factors. In some embodiments, the cytokine or growth factor measurements include measurements of 1 cytokine or growth factor. In some embodiments, the cytokine or growth factor measurements include measurements of 2 cytokines or growth factors. In some embodiments, the cytokine or growth factor measurements include measurements of 3 cytokines or growth factors. In some embodiments, the cytokine or growth factor measurements include measurements of 4 cytokines or growth factors. In some embodiments, the cytokine or growth factor measurements include measurements of 5 cytokines or growth factors. In some embodiments, the cytokine or growth factor measurements include measurements of 6 cytokines or growth factors. In some embodiments, the cytokine or growth factor measurements include measurements of 7 cytokines or growth factors. In some embodiments, the cytokine or growth factor measurements include measurements of 8 cytokines or growth factors. In some embodiments, the cytokine or growth factor measurements include measurements of 9 cytokinesMintz Docket No.062397-501001WO or growth factors. In some embodiments, the cytokine or growth factor measurements include measurements of 10 cytokines or growth factors.

[0091] In some embodiments, the cytokines or growth factors used herein include CCL-2,CTGF, FGF-2, HGF, histamine, IGF-1, IL-1β, PDGF, PDGF-BB, PF4, prostaglandin E2, TGF-β,TGF-β1, TNF, TSP-1, or VEGF. In some embodiments, the cytokines or growth factors used hereininclude a combination of CCL-2, CTGF, FGF-2, HGF, histamine, IGF-1, IL-1β, PDGF, PDGF-BB,PF4, prostaglandin E2, TGF-β, TGF-β1, TNF, TSP-1, or VEGF. In some embodiments, the cytokinesor growth factors used herein include IGF-1, CCL-2, TNF, IL-1B, PDGF-BB, EGF, CTGF, FGF-2,HGF, TSP-1, TGF-β1, PF4, or a combination thereof. In some embodiments, the cytokines or growthfactors used herein include IGF-1, CCL-2, TNF, IL-1B, PDGF-BB, EGF, CTGF, FGF-2, HGF, TSP-1, TGF-β1, and PF4. In some embodiments, the cytokines or growth factors used herein include IGF-1, CCL-2, TNF, IL-1B, PDGF-BB, FGF-2, HGF, TSP-1, TGF-β1, PF4, or a combination thereof. Insome embodiments, the cytokines or growth factors used herein include IGF-1, CCL-2, TNF, IL-1B,PDGF-BB, FGF-2, HGF, TSP-1, TGF-β1, and PF4. In some embodiments, the cytokines or growthfactors used herein include PDGF, VEGF, TNF, IGF-1, IL-1B, CTGF, or a combination thereof. In some embodiments, the cytokines or growth factors used herein include PDGF, VEGF, TNF, IGF-1, IL-1B, and CTGF. In some embodiments, the cytokines or growth factors used herein include PDGF, IGF-1, CTGF, VEGF, TGF, or a combination thereof. In some embodiments, the cytokines or growth factors used herein include PDGF, IGF-1, CTGF, VEGF, and TGF. In some embodiments, thecytokines or growth factors used herein include VEGF, PDGF, TGF-β, FGF-2, CTGF, IGF, HGF,CCL-2, PF4, prostaglandin E2, histamine, or a combination thereof. In some embodiments, thecytokines or growth factors used herein include VEGF, PDGF, TGF-β, FGF-2, CTGF, IGF, HGF,CCL-2, PF4, prostaglandin E2, and histamine. In some embodiments, the cytokines or growth factorsused herein include TNF, IL1β, IGF-1, TGF-β1, or a combination thereof. In some embodiments, thecytokines or growth factors used herein include TNF, IL1β, IGF-1, and TGF-β1. In someembodiments, the cytokines or growth factors used herein include PDGF-BB, IL-1B, HGF, TSP-1,TGF-β1, PF4, or a combination thereof. In some embodiments, the cytokines or growth factors usedherein include PDGF-BB, IL-1B, HGF, TSP-1, TGF-β1, and PF4. In some embodiments, thecytokines or growth factors used herein include CCL-2, FGF-2, HGF, TSP-1, TGF-β1, PF4, or acombination thereof. In some embodiments, the cytokines or growth factors used herein include CCL-2, FGF-2, HGF, TSP-1, TGF-β1, and PF4. In some embodiments, the cytokines or growth factorsused herein include PDGF-BB, FGF-2, HGF, TSP-1, TGF-β1, PF4, or a combination thereof. In someembodiments, the cytokines or growth factors used herein include PDGF-BB, FGF-2, HGF, TSP-1,TGF-β1, and PF4. In some embodiments, the cytokines or growth factors used herein include PDGF-BB, FGF-2, HGF, TSP-1, TGF-β1, PF4, or a combination thereof. In some embodiments, thecytokines or growth factors used herein include PDGF-BB, FGF-2, HGF, TSP-1, TGF-β1, and IGF-1.In some embodiments, the cytokines or growth factors used herein include PDGF-BB, FGF-2, HGF,Mintz Docket No.062397-501001WOTSP-1, TGF-β1, PF4, or a combination thereof. In some embodiments, the cytokines or growthfactors used herein include PDGF-BB, FGF-2, HGF, TSP-1, TGF-β1, and IGF-1. In someembodiments, the cytokines or growth factors used herein include PDGF-BB, IL-1B, HGF, TSP-1,TGF-β1, IGF-1, or a combination thereof. In some embodiments, the cytokines or growth factors usedherein include PDGF-BB, IL-1B, HGF, TSP-1, TGF-β1, and IGF-1.

[0092] Disclosed herein, in some embodiments, are predetermined cytokine or growth factorranges. In some embodiments, the predetermined cytokine or growth factor ranges are within 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, or 3 standard deviations of average cytokine or growth factor measurements in a normal population. In some embodiments, the predetermined cytokine or growth factor ranges are within 0.5 standard deviations of average cytokine or growth factor measurements in a normal population. In some embodiments, the predetermined cytokine or growth factor ranges are within 0.75 standard deviations of average cytokine or growth factor measurements in a normal population. In some embodiments, the predetermined cytokine or growth factor ranges are within 1 standard deviation of average cytokine or growth factor measurements in a normal population. In some embodiments, the predetermined cytokine or growth factor ranges are within 1.5 standard deviations of average cytokine or growth factor measurements in a normal population. In some embodiments, the predetermined cytokine or growth factor ranges are within 2 standard deviations of average cytokine or growth factor measurements in a normal population. In some embodiments, the predetermined cytokine or growth factor ranges are within 2.5 standard deviations of average cytokine or growth factor measurements in a normal population.

[0093] In some embodiments, the predetermined cytokine or growth factor ranges are withinabout 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1, about 1.2, about 1.4, about 1.6, about 1.8, about 2, about 2.2, about 2.4, about 2.6, about 2.8, or about 3 standard deviations of average cytokine or growth factor measurements in a normal, healthy, or control population. In some embodiments, the predetermined cytokine or growth factor ranges are within less than 0.4, less than 0.5, less than 0.6, less than 0.7, less than 0.8, less than 0.9, less than 1, less than 1.2, less than 1.4, less than 1.6, less than 1.8, less than 2, less than 2.2, less than 2.4, less than 2.6, less than 2.8, or less than 3 standard deviations of average cytokine or growth factor measurements in a normal, healthy, or control population. In some embodiments, the predetermined cytokine or growth factor ranges are within at least 0.4, at least 0.5, at least 0.6, at least 0.7, at least 0.8, at least 0.9, at least 1, at least 1.2, at least 1.4, at least 1.6, at least 1.8, at least 2, at least 2.2, at least 2.4, at least 2.6, at least 2.8, or at least 3 standard deviations of average cytokine or growth factor measurements in a normal, healthy, or control population.

[0094] In some embodiments, a predetermined cytokine or growth factor range of thepredetermined cytokine or growth factor ranges is at least 50 pg / mL, at least 100 pg / mL, at least 500 pg / mL, at least 1 µg / mL, at least 5 µg / mL, at least 10 µg / mL, at least 50 µg / mL, or at least 100 µg / mL, depending on the cytokine or growth factor. In some embodiments, a predetermined cytokineMintz Docket No.062397-501001WO or growth factor range of the predetermined cytokine or growth factor ranges is below 100 pg / mL, below 500 pg / mL, below 1 µg / mL, below 5 µg / mL, below 10 µg / mL, below 50 µg / mL, below 100 µg / mL, or below 500 µg / mL, depending on the cytokine or growth factor.

[0095] Some examples of amounts of cytokines or growth factors that may be useful in apredetermined cytokine or growth factor range are shown in Table 1. The amounts in the table include means and standard deviations of a general population. Table 1. Example Sentinel proteins

[0096] To obtain a cytokine or growth factor measurement, an amount may be normalized. Thenormalization may include normalizing by a housekeeping protein amount. The normalization may include normalizing by a total protein amount. The normalization may include normalizing by a platelet amount. The platelet amount may include a platelet count. The normalization may includeMintz Docket No.062397-501001WO normalizing by a platelet count. Some embodiments do not include normalization, or do not include a normalization provided here.

[0097] Normalizing may reduce variability in measurements such as Sentinel proteinmeasurements. In some cases, normalizing cytokine or growth factor amounts may result in an decrease in noise, for example a standard deviation decrease.

[0098] Normalizing may increase variability in measurements such as Sentinel proteinmeasurements. In some cases, normalizing cytokine or growth factor amounts may result in an increase in noise, for example a standard deviation increase. In some embodiments, an increase in standard deviation due to normalization (such as normalization to a platelet count) may result in measurements that are more physiologically accurate than not using the normalization.

[0099] In some embodiments, the predetermined cytokine or growth factor ranges compriseranges for each of a plurality of cytokine(s) and / or growth factor(s). In some embodiments, the cytokine or growth factor measurements are combined into a cytokine or growth factor index. A Sentinel index is an example of a combined cytokine or growth factor index. The cytokine or growth factor measurements may be combined by use of an algorithm. In some embodiments, combining the cytokine or growth factor measurements into a cytokine or growth factor index comprises obtaining an arithmetic mean, a geometric mean, a median, or a weighted average. In some embodiments, combining the cytokine or growth factor measurements into a cytokine or growth factor index comprises obtaining an arithmetic mean of cytokine or growth factor measurements. In some embodiments, combining the cytokine or growth factor measurements into a cytokine or growth factor index comprises obtaining a geometric mean of cytokine or growth factor measurements. In some embodiments, combining the cytokine or growth factor measurements into a cytokine or growth factor index comprises obtaining a median of cytokine or growth factor measurements. In some embodiments, combining the cytokine or growth factor measurements into a cytokine or growth factor index comprises obtaining a weighted average of cytokine or growth factor measurements. The algorithm may include machine learning.

[0100] In some embodiments, the predetermined cytokine or growth factor ranges comprise apredetermined cytokine or growth factor index range. Such predetermined cytokine or growth factor ranges may be useful for donor selection or platelet composition monitoring.

[0101] Sentinel proteins or other factors (e.g. cytokines or growth factors) may be measured in aplatelet-enriched sample, for example as part of a donor selection process. Sentinel proteins or other factors (e.g. cytokines or growth factors) may be measured in a pooled platelet-enriched sample, for example as part of a monitoring or quality control determination to see if a pooled platelet-enriched sample is fit for use as part of a platelet composition. The sentinel proteins or other factors may be measured in a pooled platelet-enriched sample or platelet composition in determining whether to lyophilize the pooled platelet-enriched sample or platelet composition. The sentinel proteins or other factors may be measured in a pooled platelet-enriched sample or platelet composition in determiningMintz Docket No.062397-501001WO whether to package the pooled platelet-enriched sample or platelet composition. The sentinel proteins or other factors may be measured in a pooled platelet-enriched sample or platelet composition (e.g. a reconstituted platelet composition) in determining whether to administer the pooled platelet-enriched sample or platelet composition. Platelet sample pooling

[0102] Disclosed herein, in some embodiments, are methods that include platelet sample pooling(e.g. platelet-enriched sample pooling). Platelet sample pooling may include pooling samples such as platelet-enriched samples of selected donors. Platelet sample pooling may be included in a method such as a method of making a platelet composition. Platelet sample pooling may be based on a donor selection. Pooling selected platelet concentrates is useful for reducing variability in useful biological factors such as Sentinel proteins and enhance the consistency of a drug substance such as a platelet composition.

[0103] Once donors are selected, their platelet concentrates may be pooled to create a therapeuticproduct that is consistent and optimized for use in conditions such as osteoarthritis. A pooling process may involve combining platelet concentrates from multiple donors whose profiles meet criteria, ensuring that the final product (e.g. a platelet composition comprising pooled platelet-enriched samples of selected donors) has a standardized cytokine and growth factor composition. This approach is useful for minimizing variability in biological factors such as Sentinel proteins, thereby producing a more consistent drug substance.

[0104] FIG. 2 includes an example platelet sample pooling process. Any one or a combination ofthese aspects may be used in a method herein, such as a method of making a platelet composition.

[0105] Some embodiments include pooling platelet-enriched samples. Some embodimentsinclude pooling a platelet-enriched sample with other platelet-enriched samples. The pooling may be based on a donor selection decision. The pooling may be based on donor selection decisions.

[0106] Some embodiments include not pooling a platelet-enriched sample or platelet-enrichedsamples. Some embodiments include not pooling a platelet-enriched sample with other platelet- enriched samples. Not pooling may be based on a donor selection decision. Not pooling may be based on donor selection decisions.

[0107] Some embodiments include pooling the platelet-enriched sample of the donor with otherplatelet-enriched samples when the donor age is within a predetermined age range. Some embodiments include pooling the platelet-enriched sample of the donor with other platelet-enriched samples when the donor disease status is free of a predetermined selection of diseases. Some embodiments include pooling the platelet-enriched sample of the donor with other platelet-enriched samples when the systemic inflammation measurement is below a predetermined systemic inflammation threshold. Some embodiments include pooling the platelet-enriched sample of the donor with other platelet-enriched samples when the cytokine or growth factor measurements are withinMintz Docket No.062397-501001WO predetermined cytokine or growth factor ranges. Some embodiments include pooling the platelet- enriched sample of a donor with other platelet-enriched samples when the donor age is within a predetermined age range, the donor disease status is free of a predetermined selection of diseases, the systemic inflammation measurement is below a predetermined systemic inflammation threshold, and the cytokine or growth factor measurements are within predetermined cytokine or growth factor ranges. The pooling may generate a platelet composition comprising pooled platelet-enriched samples of the group of human donors.

[0108] Some embodiments include not pooling the platelet-enriched sample of a donor withother platelet-enriched samples. Some embodiments include not pooling the platelet-enriched sample of a donor with the other platelet-enriched samples when the donor age is not within the predetermined age range. Some embodiments include not pooling the platelet-enriched sample of a donor with the other platelet-enriched samples when the donor disease status is not free of the predetermined selection of diseases. Some embodiments include not pooling the platelet-enriched sample of a donor with the other platelet-enriched samples when the systemic inflammation measurement is not below the predetermined systemic inflammation threshold. Some embodiments include not pooling the platelet-enriched sample of a donor with the other platelet-enriched samples when the cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges. A platelet composition may exclude platelet-enriched samples of any donor whose platelet-enriched sample is not pooled with the other samples.

[0109] Some embodiments include pooling the platelet-enriched sample of the donor with otherplatelet-enriched samples when the donor age is within a predetermined age range, the donor disease status is free of a predetermined selection of diseases, the systemic inflammation measurement is below a predetermined systemic inflammation threshold, and the cytokine or growth factor measurements are within predetermined cytokine or growth factor ranges, thereby generating a platelet composition comprising pooled platelet-enriched samples of the group of human donors, and not pooling the platelet-enriched sample of the donor with the other platelet-enriched samples when the donor age is not within the predetermined age range, the donor disease status is not free of the predetermined selection of diseases, the systemic inflammation measurement is not below the predetermined systemic inflammation threshold, or the cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges.

[0110] Some embodiments include pooling a number of platelet-enriched samples. For example,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more platelet-enriched samples may be pooled. In some embodiments, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, or about 20 platelet-enriched samples are pooled. In some embodiments, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, or at least 20 platelet-enrichedMintz Docket No.062397-501001WO samples are pooled. In some embodiments, less than 2, less than 3, less than 4, less than 5, less than 6, less than 7, less than 8, less than 9, less than 10, less than 11, less than 12, less than 13, less than 14, less than 15, less than 16, less than 17, less than 18, less than 19, or less than 20 platelet-enriched samples are pooled. In some embodiments, the platelet composition comprises at 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more platelet-enriched samples.

[0111] In some embodiments, pooling process involves combining platelet concentrates from atleast 3 selected donors. An optimal number of donors to pool may be at least 5 to provide consistency of a platelet composition. In some embodiments, pooling process involves combining platelet concentrates from 6 to 8 selected donors.

[0112] Some embodiments include pooling PRP samples (e.g. PRP samples of donors). In someembodiments, the pooled PRP samples are of donor subjects with systemic inflammation measurements within a systemic inflammation concentration range, and PRP samples of donor subjects with systemic inflammation measurements outside the systemic inflammation concentration range are excluded from the pooled PRP samples. In some embodiments, the pooled PRP samples are of donor subjects with CRP measurements within a CRP concentration range, and PRP samples of donor subjects with CRP measurements outside the CRP concentration range are excluded from the pooled PRP samples. In some embodiments, the pooled PRP samples are of donor subjects free of any autoimmune diseases, cancer, osteoarthritis, and cardiovascular disease, and PRP samples of donor subjects who have the autoimmune disease, cancer, osteoarthritis, or cardiovascular disease are excluded from the pooled PRP samples. In some embodiments, the pooled PRP samples are of donor subjects with one or more cytokine or growth factor measurements in a preselected cytokine or growth factor concentration range, and PRP samples of donor subjects with one or more cytokine or growth factor measurements outside the preselected cytokine or growth factor concentration range are excluded from the pooled PRP samples. In some embodiments, the pooled PRP samples comprise at least 3 PRP samples. In some embodiments, the pooled PRP samples comprise 6-8 PRP samples.

[0113] In some embodiments, pooled donor samples include or consist of male donor samples. Insome embodiments, pooled donor samples include or consist of female donor samples. In some embodiments, pooled donor samples include a combination of male and female donor samples. Using platelet compositions

[0114] Disclosed herein, in some embodiments, are methods of using a platelet composition. Theuse may include administration. The use may include treatment. The treatment may be for a disease or disorder, or may be cosmetic. The treatment may include another treatment such as surgery or administration of another drug in addition to the administration of a platelet composition.

[0115] The treatment may be for a disease or disorder. Some embodiments include administeringthe platelet composition to a subject in need of treatment for a disease or disorder. In some embodiments, the disease comprises an autoimmune disease, an inflammatory disease, a degenerativeMintz Docket No.062397-501001WO joint disease. The disease may include an autoimmune disease. The disease may include an inflammatory disease. The disease may include a degenerative disease. The degenerative disease may be a degenerative joint disease. The disease may include a degenerative joint disease.

[0116] In some embodiments, the disease or disorder comprises osteoarthritis, muscle strain,tendinopathy, tendinosis, or a muscle-fascial injury. The disease or disorder may include osteoarthritis. The disorder may include a muscle strain. The disease or disorder may include tendinopathy. The disease or disorder may include tendinosis. The disorder may include an injury. The disorder may include a muscle-fascial injury. The disease or disorder may include a muscle disease or disorder. The disease or disorder may include a fascial disease or disorder.

[0117] In some embodiments, the treatment comprises a dermatological, orthopedic, sportsmedicine, cosmetic surgery, cosmeceutical, wound healing, dry eyes (e.g. dry eye disease), alopecia, ophthalmic surgery, neurosurgery, or general surgery treatment. The disease or disorder may be dermatological. The disease or disorder may be orthopedic. Examples of orthopedic diseases or disorders may include lateral epicondylitis, plantar fasciitis, anterior cruciate ligament (ACL) repair, rotator cuff tendinopathy, osteoarthritis (OA, e.g. OA of a hip or knee), high ankle sprains, rotator cuff repair, patellar and Achilles tendinopathies, hamstring injuries, or medial epicondylitis. The disease or disorder may be sports related. The disease or disorder may include a wound. The disease or disorder may include dry eye disease. The disease or disorder may include alopecia. The disease or disorder may be ophthalmic. A concurrent treatment may involve surgery such as ophthalmic surgery, neurosurgery, or general surgery. The disease or disorder may be neurological. The disease or disorder may be general.

[0118] The treatment may be cosmetic. Some embodiments include administering the plateletcomposition to a subject as a cosmetic treatment. The treatment may include a cosmetic surgery. The treatment may include a cosmeceutical.

[0119] In some embodiments, the administration comprises an injection. In some embodiments,the administration is to a joint, ankle, elbow, wrist, neck, spine, lower back. For example, a treatment may include an injection to a joint, ankle, elbow, wrist, neck, spine, lower back. An administration may be made to a joint. An administration may be made to an ankle. An administration may be made to an elbow. An administration may be made to a wrist. An administration may be made to a neck. An administration may be made to a spine. An administration may be made to a lower back. In some embodiments, the administration is to an area of poor vascularization.

[0120] In some embodiments, the injection is intraarticular, intramuscular (e.g. for hamstringinjuries), subdermal injection, or subcutaneous. An administration or injection may be intraarticular. An administration or injection may be intramuscular. An administration or injection may be subdermal. An administration or injection may be subcutaneous. In some embodiments, the injection comprises microneedling.Mintz Docket No.062397-501001WO

[0121] In some embodiments, the administration is to an eye. In some embodiments, the plateletcomposition includes an eye drop. In some embodiments, the administration is to an eye as an eye drop.

[0122] A treatment or administration may have an effect, for example an improvement of asymptom of a disease or disorder. A treatment or administration may have an effect on a disease. A treatment or administration may have an effect on a disorder. A treatment or administration may have a cosmetic effect. An effect may be beneficial. An effect may be an improvement. An effect may be on a symptom. The effect may be relative to a baseline before said treatment. The treatment may affect a measurement of an effect relative to a baseline measurement. The treatment may result in an improvement in OA or a symptom of OA, relative to before treatment.

[0123] In some embodiment, the treatment affects a cell or tissue of a subject. In someembodiment, the treatment affects a cell. In some embodiment, the treatment affects a tissue.

[0124] In some embodiments, the subject in need of treatment is a human subject, or the subjectadministered a cosmetic product is a human subject. A treated subject may be a vertebrate. A treated subject may be an animal. A treated subject may be a mammal. A treated subject may be a primate. A treated subject may be a human. Kits

[0125] Disclosed herein, in some embodiments, are kits. The kit may include a plateletcomposition. The platelet composition may be liquid or may be lyophilized. The kit may include a buffer. The kit may include a reconstitution buffer. The kit may include instructions for use in a method of use, such as a method of treatment. Definitions

[0126] Unless defined otherwise, all terms of art, notations and other technical and scientificterms or terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art.

[0127] Throughout this application, various embodiments may be presented in a range format. Itshould 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 disclosure. 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 1Mintz Docket No.062397-501001WO 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.

[0128] As used in the specification and claims, the singular forms “a”, “an” and “the” includeplural references unless the context clearly dictates otherwise. For example, the term “a sample”includes a plurality of samples, including mixtures thereof.

[0129] The terms “determining,” “measuring,” “evaluating,” “assessing,” “assaying,” and“analyzing” are often used interchangeably herein to refer to forms of measurement. The termsinclude determining if an element is present or not (for example, detection). These terms can include quantitative, qualitative or quantitative and qualitative determinations. Assessing can be relative orabsolute. “Detecting the presence of” can include determining the amount of something present inaddition to determining whether it is present or absent depending on the context.

[0130] The terms “subject,” “individual,” or “patient” are often used interchangeably herein. A“subject” can be a biological entity containing expressed genetic materials. The biological entity canbe a plant, animal, or microorganism, including, for example, bacteria, viruses, fungi, and protozoa. The subject can be tissues, cells and their progeny of a biological entity obtained in vivo or cultured in vitro. The subject can be a mammal. The mammal can be a human. The subject may be diagnosed or suspected of being at high risk for a disease. In some cases, the subject is not necessarily diagnosed or suspected of being at high risk for the disease.

[0131] As used herein, the term “about” a number refers to that number plus or minus 15% ofthat number. The term “about” a range refers to that range minus 15% of its lowest value and plus15% of its greatest value.

[0132] As used herein, the terms “treatment” or “treating” are used in reference to apharmaceutical or other intervention regimen for obtaining beneficial or desired results in the recipient. Beneficial or desired results include but are not limited to a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit may refer to eradication or amelioration of symptoms or of an underlying disorder being treated. Also, a therapeutic benefit can be achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. A prophylactic effect includes delaying, preventing, or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof. For prophylactic benefit, a subject at risk of developing a particular disease, or to a subject reporting one or more of the physiological symptoms of a disease may undergo treatment, even though a diagnosis of this disease may not have been made.

[0133] The section headings used herein are for organizational purposes only and are not to beconstrued as limiting the subject matter described.Mintz Docket No.062397-501001WO EXAMPLES

[0134] The following examples are included for illustrative purposes only and are not intended tolimit the scope of the invention. Example 1: platelet donor selection and pooling

[0135] A group of donor PRP samples was screened for platelet cytokine and growth factorlevels, including Sentinel proteins. Donors whose profiles met the thresholds were selected, and their platelet concentrates were pooled in silico to simulate creation of a standardized therapeutic product. An analysis was conducted to assess the variability in the concentration of useful biological factors. The results showed a significant reduction in variability, demonstrating the effectiveness of the pooling method in producing a more consistent drug substance. Methods

[0136] Sentinel index calculation: a Sentinel index was calculated for each sample bystandardizing the concentrations of 6 Sentinel proteins and summing the standardized values. Samples of subjects with CRP levels above 3.0 mg / L were excluded to focus on more consistent profiles. Samples of subjects over 40 and under 18 years of age were excluded or were not provided. Samples of subjects who self-identified or were tested as positive for a group of diseases were excluded or were not provided. The group of diseases included infectious disease, cancer, autoimmune disease, and cardiovascular disease.

[0137] Simulation of pooling process: a computational pooling was performed, assuming Nequal amounts of 1 / N volumes from each donor such that a single pooled unit was achieved mathematically with each donations comprising 1 / Nth of the sample volume.

[0138] Pool sizes ranging from 2 to 10 samples were tested. For each pool size, 1,000simulations were performed. In each simulation, a random subset of samples was selected with replacement, and the mean Sentinel index for the subset was calculated. The standard deviation of the pooled Sentinel index values was computed for each pool size, providing a measure of variability.

[0139] Rate of change in variability: to identify a point of diminishing returns, the rate ofchange in standard deviation was calculated as a difference in variability between successive pool sizes. This analysis aimed to pinpoint where increasing the pool size no longer resulted in significant reductions in variability. Results

[0140] Pooling platelet donations is a useful strategy to achieve consistent proteinconcentrations, particularly for Sentinel proteins that are useful in therapeutic applications such as those provided herein. A diminishing returns analysis was conducted to identify the optimal pool sizeMintz Docket No.062397-501001WO where the reduction in variability of the Sentinel Index slows, indicating minimal additional benefit from pooling more samples.

[0141] The analysis revealed a decreasing trend in the standard deviation of the Sentinel index asthe pool size increased, with the most substantial reductions occurring at smaller pool sizes (FIG.3A- 3B). The rate of change in standard deviation was most pronounced between pool sizes of 2 to 4 samples, indicating a significant reduction in variability. However, the rate of change decreased notably around pool sizes of 6 to 7 samples, where the reduction in variability slowed considerably. Beyond this point, the standard deviation continued to decrease, but at a much slower rate, indicating diminishing returns. Conclusion

[0142] The Diminishing Returns Analysis suggests that pooling approximately 6 to 7 plateletdonations optimizes the consistency of the Sentinel index. At this pool size, the reduction in variability is substantial, while further increases in pool size yield minimal additional benefits. This finding provides a guideline for determining the optimal number of samples to pool, balancing the need for consistency with cost considerations. This study exemplifies the usefulness of selection processes herein and pooling of samples as described. Example 2

[0143] A study was performed, which included the following:Sentinel protein selection: several platelet proteins were identified that may be useful in treating knee OA with platelet rich plasma (PRP). Sample collection and characterization: collect platelet donations from 60 adults ranging in age from 18 to 75, measure platelet count, total protein concentration, and inflammatory status. Protein array studies: perform protein array binding studies to determine the concentrations of 6 Sentinels and 26 additional proteins. Data analysis: analyze data to determine relationships amongst protein concentrations, age, sex, and systemic inflammation status (here, CRP status). Create Inflammatory Index and Sentinel index. Insight generation: identify donor selection criteria based on age, inflammatory status, and Sentinel read-outs. Identify appropriate number of donor platelet units to combine that balances Sentinel factor variability to improve consistent platelet compositions. Findings

[0144] Normalization by platelet count: normalizing protein concentrations by platelet countdid not reduce the variability of Sentinel proteins. Instead, the standard deviations increased forMintz Docket No.062397-501001WO Sentinel proteins after normalization. Therefore, measurements of Sentinel proteins may not need to be normalized by platelet count for use in donor selection or monitoring of a platelet composition.

[0145] Sentinel protein measurements: Concentrations for Sentinel proteins were measured inpicograms per milliliter (pg / ml) or nanograms per milliliter (ng / ml). Comparative Gaussian distribution curves were generated for the 6 Sentinel proteins used in this example, and differences in concentration profiles between healthy samples and a full dataset were identified. For example, plots were provided that represented a probability density function of Sentinel protein concentrations, where curves depicted Gaussian distributions, calculated using the mean and standard deviation for healthy samples (age < 40, CRP < 3.0 mg / L) and for all samples. Plots included mean concentrations for each distribution. The plots were used to identify shifts in concentration distributions and central tendencies between the two populations, providing insights into the variability and consistency of Sentinel protein levels in relation to donor health and demographic factors.

[0146] Correlation with Age and CRP: PDGF-BB showed the strongest positive correlationwith age (0.416) compared to other Sentinel proteins. G-CSF, follistatin, HG-EGF, PLGF, angiopoietin-2 and fibrinogen were also strongly correlated with age, having a correlation coefficient of at least about 0.3

[0147] Most proteins exhibited positive correlations with age, including HGF and FGF-2, whileTSP-1, PF4, and TGF-β1 showed negative correlations. HGF and VEGF-D had weak correlations ordid not correlate with age in the study.

[0148] Proteins strongly correlated with high plasma CRP levels above 3.0 mg / L included leptin,AGP, haptoglobin, SAP, EGF, Thrombospondin-1, CRP, IL-8, L-Selectin, PF4, epidermal growth factor, and IL-8. Some such proteins may serve as biomarkers for youthful or healthy states. L-selectin, VEGF-A and TGF-β1 had weak correlations or did not correlate with high CRP in the study.

[0149] Inflammatory Index and Age Cutoff: an Inflammatory Index was developed using 11proteins highly correlated with CRP. As shown in FIG. 4A, he Inflammatory Index increased withage, suggesting a donor age cutoff of 60 years, or preferably 40 years, to minimize inflammatory profiles. Data stratifying inflammatory indexes for men and women separately are also included in FIG.4B.

[0150] Sentinel Index and Optimal Pool Size: a Sentinel Index was created by aggregating thestandardized values of the 6 Sentinel proteins. Variability analysis showed that pooling approximately6 platelet donations optimizes the consistency of the Sentinel Index, achieving a significant reduction in variability while avoiding diminishing returns. Conclusions

[0151] This study showed a usefulness of using Sentinel proteins in making a consistent anduseful platelet composition. For example, using Sentinel proteins in donor selection or in monitoring a platelet composition may help ensure a platelet composition is consistent and useful for treatment.Mintz Docket No.062397-501001WO

[0152] This study showed that eliminating donors with high systemic inflammation markerlevels (e.g. CRP over 3.0 mg / L) is useful in creating a platelet composition. This study showed that eliminating donors under an age threshold such as 40 or 60 is useful for minimizing inflammatory profiles within platelet compositions. This study showed that adopting a platelet pooling strategy (for example, that utilizes approximately 6 donations) may optimize consistency of the Sentinel Index. ***

[0153] While preferred embodiments of the present invention have been shown and describedherein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

Mintz Docket No.062397-501001WO CLAIMS 1. A method of making a platelet composition, comprising:obtaining donor information for a group of human donors; optionally obtaining biofluid samples of the group of human donors; obtaining platelet-enriched samples of the group of human donors; and for a donor of the group of donors: determining an age of the donor from the donor information, determining a disease status of the donor from the donor information or from disease status measurements obtained from one or more of the biofluid samples, obtaining a systemic inflammation measurement for the donor from the donor information or from a systemic inflammation measurement obtained from one or more of the biofluid samples, obtaining cytokine or growth factor measurements for the donor from a platelet- enriched sample of the donor, pooling the platelet-enriched sample of the donor with other platelet-enriched samples when the donor age is within a predetermined age range, the donor disease status is free of a predetermined selection of diseases, the systemic inflammation measurement is below a predetermined systemic inflammation threshold, and the cytokine or growth factor measurements are within predetermined cytokine or growth factor ranges, thereby generating a platelet composition comprising pooled platelet-enriched samples of the group of human donors, and not pooling the platelet-enriched sample of the donor with the other platelet-enriched samples when the donor age is not within the predetermined age range, the donor disease status is not free of the predetermined selection of diseases, the systemic inflammation measurement is not below the predetermined systemic inflammation threshold, or the cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges.

2. The method of claim 1, wherein the platelet-enriched samples are generated via aseries of centrifugation steps or by apheresis of a blood sample.

3. The method of claim 1, wherein the biofluid sample is a whole blood sample or aplasma sample.

4. The method of claim 1, wherein obtaining the platelet-enriched sample of the donor isoptional, and wherein: the platelet-enriched sample of the donor is obtained when the donor age is within the predetermined age range, the donor disease status is free of the predetermined selection of diseases, and the systemic inflammation measurement is below the predetermined systemic inflammation threshold; andMintz Docket No.062397-501001WO the platelet-enriched sample of the donor is not obtained when the donor age is not within the predetermined age range, the donor disease status is not free the predetermined selection of diseases, or the systemic inflammation measurement is not below the predetermined systemic inflammation threshold, wherein when the platelet-enriched sample of the donor is not obtained, no platelet- enriched sample of the donor is pooled with the other platelet-enriched samples.

5. The method of claim 1, wherein the predetermined age range includes an age of atleast 10 years, at least 12 years, at least 14 years, at least 16 years, at least 18 years, at least 20 years, at least 22 years, at least 24 years, at least 26 years, at least 28 years, or at least 30 years.

6. The method of claim 1, wherein the predetermined age range includes an age below60 years, below 58 years, below 56 years, below 54 years, below 52 years, below 50 years, below 48 years, below 46 years, below 44 years, below 42 years, below 40 years, below 38 years, below 36 years, below 34 years, or below 32 years, or below 30 years.

7. The method of claim 1, wherein the predetermined selection of diseases comprisesinfectious disease, cancer, autoimmune disease, and cardiovascular disease.

8. The method of claim 1, wherein the predetermined selection of diseases compriseshuman immunodeficiency virus, human T-cell lymphotropic virus, hepatitis B, hepatitis C, syphilis, West Nile virus, and Chagas disease.

9. The method of claim 1, wherein the systemic inflammation measurement comprises aC-reactive protein (CRP) measurement, and the predetermined systemic inflammation threshold comprises a predetermined CRP threshold.

10. The method of claim 9, wherein the predetermined CRP threshold is 0 mg / L (e.g.undetectable), 0.5 mg / L, 1 mg / L, 1.5 mg / L, 2 mg / L, 2.5 mg / L, 3 mg / L, 3.5 mg / L, 4 mg / L, 4.5 mg / L, 5 mg / L, 5.5 mg / L, 6 mg / L, 6.5 mg / L, 7 mg / L, 7.5 mg / L, 8 mg / L, 8.5 mg / L, 9 mg / L, 9.5 mg / L, or 10 mg / L.

11. The method of claim 1, wherein the systemic inflammation measurement comprisesan erythrocyte sedimentation rate (ESR) measurement, and the predetermined systemic inflammation threshold comprises a predetermined ESR threshold.

12. The method of claim 11, wherein the predetermined ESR threshold is 10 mm / hour, 15mm / hour, 20 mm / hour, 25 mm / hour, 30 mm / hour, 35 mm / hour, or 40 mm / hour.

13. The method of claim 1, wherein the predetermined cytokine or growth factor rangesare within 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, or 3 standard deviations of average cytokine or growth factor measurements in a normal population.

14. The method of claim 1, wherein a predetermined cytokine or growth factor range ofthe predetermined cytokine or growth factor ranges is at least 50 pg / mL, at least 100 pg / mL, at least 500 pg / mL, at least 1 µg / mL, at least 5 µg / mL, at least 10 µg / mL, at least 50 µg / mL, or at least 100 µg / mL.Mintz Docket No.062397-501001WO 15. The method of claim 1, wherein a predetermined cytokine or growth factor range ofthe predetermined cytokine or growth factor ranges is below 100 pg / mL, below 500 pg / mL, below 1 µg / mL, below 5 µg / mL, below 10 µg / mL, below 50 µg / mL, below 100 µg / mL, or below 500 µg / mL.

16. The method of claim 1, wherein the predetermined cytokine or growth factor rangescomprise ranges for each of a plurality of cytokine(s) and / or growth factor(s).

17. The method of claim 1, wherein the cytokine or growth factor measurements arecombined into a cytokine or growth factor index.

18. The method of claim 17, wherein combining the cytokine or growth factormeasurements into a cytokine or growth factor index comprises obtaining an arithmetic mean, a geometric mean, a median, or a weighted average.

19. The method of claim 1, wherein the predetermined cytokine or growth factor rangescomprise a predetermined cytokine or growth factor index range.

20. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a chemokine C-C motif ligand 2 (CCL-2) measurement.

21. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a connective tissue growth factor (CTGF) measurement.

22. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a fibroblast growth factor (FGF) measurement.

23. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a fibroblast growth factor 2 (FGF-2) measurement.

24. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a hepatocyte growth factor (HGF) measurement.

25. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a histamine measurement.

26. The method of claim 1, wherein the cytokine or growth factor measurementscomprise an insulin-like growth factor 1 (IGF-1) measurement.

27. The method of claim 1, wherein the cytokine or growth factor measurementscomprise an interleukin-1 beta (IL-1β) measurement.

28. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a platelet-derived growth factor (PDGF) measurement.

29. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a platelet-derived growth factor BB (PDGF-BB) measurement.

30. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a platelet factor 4 (PF4) measurement.

31. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a prostaglandin E2 measurement.Mintz Docket No.062397-501001WO 32. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a transforming growth factor-beta (TGF-β) measurement.

33. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a transforming growth factor-beta 1 (TGF-β1) measurement.

34. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a tumor necrosis factor (TNF) measurement.

35. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a thrombospondin-1 (TSP-1) measurement.

36. The method of claim 1, wherein the cytokine or growth factor measurementscomprise an epidermal growth factor (EGF) measurement.

37. The method of claim 1, wherein the cytokine or growth factor measurementscomprise a vascular endothelial growth factor (VEGF) measurement.

38. The method of claim 1, further comprising enriching the platelet composition furtherby centrifuging the pooled platelet-enriched samples and removing erythrocytes and leukocytes.

39. The method of claim 1, further comprising lyophilizing the platelet composition.

40. The method of claim 39, further comprising obtaining second cytokine or growthfactor measurements from the platelet composition before the lyophilization.

41. The method of claim 1, further comprising lyophilizing the platelet composition whenthe second cytokine or growth factor measurements are within the predetermined cytokine or growth factor ranges, and not lyophilizing the platelet composition when the second cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges.

42. The method of claim 39, further comprising reconstituting the lyophilized plateletcomposition.

43. The method of claim 1, further comprising combining the platelet composition with abuffer.

44. The method of claim 1, further comprising combining the platelet composition with amonosaccharide, disaccharide, or polysaccharide.

45. The method of claim 1, further comprising formulating the platelet composition into apharmaceutical composition.

46. The method of claim 45, wherein the pharmaceutical composition comprises a liquidsolution.

47. The method of claim 45, wherein the pharmaceutical composition comprises alyophilized platelet composition.

48. The method of claim 47, wherein the lyophilized platelet composition is formulatedfor reconstitution with a reconstitution buffer by a medical practitioner.

49. The method of claim 1, further packaging the platelet composition.Mintz Docket No.062397-501001WO 50. The method of claim 49, further comprising obtaining second or third cytokine orgrowth factor measurements from the platelet composition before the packaging.

51. The method of claim 1, further comprising packaging the platelet composition whenthe second or third cytokine or growth factor measurements are within the predetermined cytokine or growth factor ranges, and not packaging the platelet composition when the second or third cytokine or growth factor measurements are not within the predetermined cytokine or growth factor ranges.

52. The method of claim 45, further comprising administering the platelet composition toa subject in need of treatment for a disease or disorder.

53. The method of claim 52, wherein the disease comprises an autoimmune disease, aninflammatory disease, a degenerative joint disease.

54. The method of claim 52, wherein the disease or disorder comprises osteoarthritis,muscle strain, tendinopathy, tendinosis, or a muscle-fascial injury.

55. The method of claim 1, further comprising formulating the platelet composition into acosmetic composition.

56. The method of claim 55, further comprising administering the platelet composition toa subject as a cosmetic treatment.

57. The method of claim 52, wherein the treatment comprises a dermatological,orthopedic, sports medicine, cosmetic surgery, cosmeceutical, wound healing, dry eye disease, alopecia, ophthalmic surgery, neurosurgery, or general surgery treatment.

58. The method of claim 52, wherein the administration comprises an injection.

59. The method of claim 52, wherein the administration is to a joint, ankle, elbow, wrist,neck, spine, lower back.

60. The method of claim 52, wherein the administration is to an area of poorvascularization in the subject.

61. The method of claim 60, wherein the injection is intraarticular, intramuscular (e.g. forhamstring injuries), subdermal injection, or subcutaneous.

62. The method of claim 60, wherein the injection comprises microneedling.

63. The method of claim 52, wherein the administration is to an eye as an eye drop.

64. The method of claim 1, wherein the biofluid samples comprise blood samples, serumsamples, plasma samples, or a combination thereof.

65. The method of claim 1, wherein the platelet-enriched samples are enriched forplatelets by about 1.5x, about 2x, about 2.5x, about 3x, about 3.5x, about 4x, about 4.5x, about 5x, about 5.5x, about 6x, about 6.5x, about 7x, about 7.5x, about 8x, about 8.5x, about 9x, about 9.5x, about 10x, or a range of any two of the aforementioned values, relative to a non-enriched plasma or blood sample.

66. The method of claim 1, wherein the platelet-enriched samples are enriched forplatelets by about 2-6 times relative to a non-enriched plasma or blood sample.Mintz Docket No.062397-501001WO 67. The method of claim 1, wherein the platelet composition comprises at 2, 3, 4, 5, 6, 7,, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more platelet-enriched samples.

68. A platelet composition, generated using the method of claim 1.

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