Extracellular vesicle composition for the treatment of graft-versus-host disease

Abstract

A method for treating organ transplant recipients by administering an extracellular vesicle composition has been disclosed.

Description

[0001] Cross-referencing This application claims the benefit of U.S. Provisional Patent Application No. 63 / 596,952, filed November 7, 2023, and U.S. Provisional Patent Application No. 63 / 712,072, filed October 25, 2024, each of which is incorporated herein by reference in its entirety. Background Technology

[0002] Over the past decade, the use of mesenchymal stem cells (MSCs) in the treatment of severe inflammatory conditions has increased, including graft-versus-host disease (GvHD), systemic lupus erythematosus, myocardial infarction, multiple sclerosis, Crohn's disease, and rejection of solid abdominal organs. These studies highlight the promising therapeutic benefits of the anti-inflammatory and immunomodulatory properties of MSCs. MSCs have been shown to suppress T cell proliferation in response to allogeneic antigens and mitotic stimulation, while increasing the regulatory T cell (Treg) population, which is useful for the treatment of acute cell rejection. Particularly for GvHD, MSCs have demonstrated driving T cell-mediated immunosuppressive activity in GvHD-specific animal models and have been successfully used to treat GvHD in both pediatric and adult patient populations.

[0003] Unfortunately, cell therapy has several logistical limitations, including product scalability and reproducibility, temperature control during transport, the need for cryopreservation, alloimmune response, pulmonary retention when using intravenous delivery, and access to product sources. Summary of the Invention

[0004] In some respects, this article provides a method for treating graft-versus-host disease in subjects in need, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs); wherein at least 80% of the EVs are CD63+, CD9-, or CD81-.

[0005] In some respects, this article provides a method for treating graft-versus-host disease in a subject in need, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs); wherein the one or more EVs comprise hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-21-5p, hsa-miR-221-3p, hsa-miR-222-3p, hsa-miR-2 The miRNA sequences are 2-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, or hsa-miR-940, or combinations of two or more thereof. The miRNA sequences are available at https: / / www.mirbase.org / .

[0006] In some respects, this article provides a method for treating graft-versus-host disease in subjects in need, comprising administering to the subject a composition comprising: ferritin, IGFBP-4 (insulin-like growth factor binding protein-4), IL-1 R6 (interleukin-1 receptor 6), LAMP2 (lysosome-associated membrane glycoprotein 2), bIG-H3 (transforming growth factor-β-inducible protein ig-h3), GPR115 (adhesion G protein-coupled receptor F4), CD63 antigen, CD109 antigen, serine protease inhibitor F1 (serpin F1) (pigment epithelium-derived factor), IGFBP-6 (insulin-like growth factor binding protein-6), HS3ST4 (heparin sulfate glucosamine 3-O-sulfotransferase 4), OPN (osteopontin), PAI-1 (plasminogen activator inhibitor-1 or SERPINE 1), and cathepsin B. B), IGFBP-2 (insulin-like growth factor binding protein-2), Semaphorin 6C, IGF-2 (insulin-like growth factor-2), Sortilin, serine protease inhibitor B6, Dkk-3 (Dickkopf-related protein 3), CNTF (ciliary neurotrophic factor), TSP-1 (thrombospondin 1), GM-CSF Ra (granulocyte-macrophage colony-stimulating factor receptor subunit α), Thrombomomodulin, Endoglycan, Podocalyxin-like protein 2, IGFBP-3 (insulin-like binding protein-3), RGM-C (hemojuvelin), PF4 (platelet factor 4), MIF (macrophage migration inhibitory factor), TGM4 (protein-glutamine-γ-glutamyltransferase 4), Periostin, Furin, TIMP-1 (MMP tissue inhibitor 1), Decorin, PCK1 (cytosol-type phosphoenolpyruvate carboxykinase). The following are considered as a combination of two or more of the following: cytosolic, CD9 antigen, CD99 antigen, CA2 (carbonic anhydrase 2), PRDX4 (peroxidoreductase-4), transferrin, DcR3 (tumor necrosis factor receptor superfamily member 6B), GP73 (Golgi membrane protein 1), CD81 antigen, lumican, or TIMP-2 (MMP tissue inhibitor 2).

[0007] In some implementations, one or more EVs include hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-21-5p, hsa-miR-221-3p, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, and hsa-miR-2 3b-3p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, or hsa-miR-940, or a combination of two or more thereof.In some embodiments, the composition comprises ferritin, IGFBP-4 (insulin-like growth factor binding protein-4), IL-1 R6 (interleukin-1 receptor 6), LAMP2 (lysosome-associated membrane glycoprotein 2), bIG-H3 (transforming growth factor-β-inducible protein ig-h3), GPR115 (adhesion G protein-coupled receptor F4), CD63 antigen, CD109 antigen, serine protease inhibitor F1 (pigment epithelial-derived factor), IGFBP-6 (insulin-like growth factor binding protein-6), HS3ST4 (heparanol sulfate glucosamine 3-O-sulfotransferase 4), OPN (osteopontin), and PAI-1 (plasminogen activator inhibitor-1 or sERPINE). 1) Cathepsin B, IGFBP-2 (insulin-like growth factor binding protein-2), brain signaling protein 6C, IGF-2 (insulin-like growth factor-2), sorting protein, serine protease inhibitor B6, Dkk-3 (Dickkopf-related protein 3), CNTF (ciliary neurotrophic factor), TSP-1 (thromboretin-1), GM-CSF Ra (granulocyte-macrophage colony-stimulating factor receptor subunit α), coagulation regulatory protein, endosaccharide, (podocyte protein-like protein 2) IGFBP-3 (insulin-like binding protein-3), RGM-C (hepcidin regulatory protein), PF4 (platelet factor 4), MIF (macrophage migration inhibitory factor), TGM4 (protein-glutamine-γ-glutamyltransferase 4), periosteal protein, furin protease, TIMP-1 (MMP tissue inhibitor 1), gypsum proteoglycan, PCK1 (cytosol-type phosphoenolpyruvate carboxykinase), CD9 antigen, CD99 antigen, CA2 (carbonic anhydrase 2), PRDX4 (peroxide reductase-4), transferrin, DcR3 (tumor necrosis factor receptor superfamily member 6B), GP73 (Golgi membrane protein 1), CD81 antigen, luminescent proteoglycan, or TIMP-2 (MMP tissue inhibitor 2), or a combination of two or more thereof.

[0008] In some implementations, the subject has received an organ transplant or allogeneic graft. In some implementations, the subject has received a modified multi-organ allogeneic graft, a single intestinal allogeneic graft, or a single liver allogeneic graft. In some implementations, the subject exhibits one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection. In some implementations, the subject exhibits one or more signs or symptoms selected from the following: crypt cell apoptosis, crypt loss, ulceration, allogeneic graft cell rejection, bile duct injury, portal vein endotheliitis, centrizonal hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, or combinations thereof.

[0009] In some implementations, the subject is receiving corticosteroids. In some implementations, the subject is receiving high-dose corticosteroids. In some implementations, the subject experiences improvement in one or more of the following signs or symptoms after treatment: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, pruritus, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, or combinations thereof.

[0010] In some embodiments, the dose of the therapeutic MSC secretome composition administered to the subject is a cell equivalent dose of 700,000 to 7,000,000 cells / kg. In some embodiments, administration includes intravenous administration. The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution may be about 100 mL. The number of EVs in the composition may be about 10 million to about 1 trillion. The number of EVs in the composition may be about 1 trillion. The number of EVs in the composition may be about 10 billion to about 1 trillion.

[0011] In some embodiments, the composition is prepared by a method comprising the following steps: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce MSC conditioned medium: (i) oxygen tension below 5%; and (ii) a medium with pH below 7; (b) harvesting the MSC conditioned medium; and (c) preparing the MSC conditioned medium to produce the composition.

[0012] In some embodiments, the method includes preparing the composition prior to application, wherein the preparation of the composition is carried out by a method comprising the following steps: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under conditions of (i) oxygen tension below 5%; and (ii) a medium with a pH below 7; (b) harvesting the MSC conditioned medium; and (c) preparing the MSC conditioned medium to produce the composition.

[0013] In some embodiments, the culture medium is serum-free. In some embodiments, the culture medium has a glucose concentration of less than 4.5 g / L. In some embodiments, preparing MSC conditioned medium involves replacing the conditioned medium with a pharmaceutically acceptable formulation. In some embodiments, the pharmaceutically acceptable formulation comprises saline. In some embodiments, the composition contains at least 6 × 10⁶ g / L of glucose. 10 Up to 8 × 10 10 The composition contains extracellular vesicles (EVs) per ml and is administered in doses of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml. The composition may have an EV concentration of at least about 60 billion EVs per ml, for example, about 60 billion to about 250 billion EVs per ml, or about 60 billion to about 80 billion EVs per ml. Optionally, about 5, 10, 15, or 20 ml is administered. As a non-limiting example, about 5, 10, 15, or 20 ml of the composition is diluted in an intravenous solution (such as saline) and administered to a subject. The intravenous solution may be about 100 ml. The number of EVs in the composition may be about 10 million to about 1 trillion. The number of EVs in the composition may be about 1 trillion. The number of EVs in the composition may be about 10 billion to about 1 trillion.

[0014] In some embodiments, this document discloses a method for treating patients who have undergone organ transplantation by administering an extracellular vesicle composition derived from bone marrow mesenchymal stem cells. One aspect of this disclosure is a method for treating graft-versus-host disease in a subject, comprising administering to the subject a composition containing a therapeutic mesenchymal stem cell (MSC) secretome composition prepared by a method comprising the steps of: (a) culturing bone marrow-derived MSCs under conditions of: (i) oxygen tension below 5%; and (ii) a medium with a pH below 7; (b) harvesting the MSC conditioned medium; and (c) preparing the MSC conditioned medium to produce the therapeutic MSC secretome composition, wherein the therapeutic MSC secretome composition comprises proteins and extracellular vesicles produced by the bone marrow-derived MSCs in step (a). In some embodiments, the subject has received an organ transplant or an allogeneic graft. In some embodiments, the subject has received a modified multi-organ allogeneic graft, a single intestinal allogeneic graft, or a single liver allogeneic graft. In some implementations, the subject exhibits one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection. In some implementations, the subject exhibits one or more signs or symptoms selected from the following: crypt cell apoptosis, crypt loss, ulceration, allogeneic graft cell rejection, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof. In some implementations, the subject is receiving corticosteroids. In some implementations, the subject is receiving high-dose corticosteroids.

[0015] In some embodiments, the subject experiences improvement in one or more of the following signs or symptoms after treatment: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, pruritus, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, or combinations thereof. In some embodiments, the therapeutic MSC secretome composition is administered to the subject at a cell equivalent dose of 700,000 to 7,000,000 cells / kg. In some embodiments, the culture medium is serum-free. In some embodiments, the culture medium has a glucose concentration of less than 4.5 g / L. In some embodiments, at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63+CD9-CD81-. In some embodiments, the therapeutic MSC secretome composition comprises one or more of the following proteins: ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1 (Serpin F1), OPN, PAI-1, DAPP1, cathepsin B, semaphorin 6C, PDGF Rα, sorting protein (Sortilin), and serine protease inhibitor B6 (Serpin F1). B6), Dkk-3, Thrombomomodulin, PF4, MIF, Periodin, Furin, TIMP-1, Decorin, PCK1, CD99, CD63, CD9, CD81, Transferrin, DcR3, Lumican, TIMP-2, SLITRK5, FAP, Artemin, DPPII, cIAP-1, Pentraxin 3, Visfatin, Neprilysin, Albumin, Galectin-1, UNC5H3, IL-20 Rβ, SREC-II, JAM-C, TNF-α, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, Plexin A4, EMMPRIN, p53, Semaphorin 7A, NKp80, Cystatin B, OsteoadherinMidkine, Calreticulin, Osteoactivin, Legumin, TAZ, Catepsin L, RBP4, Serpin A4, JAM-A, MCSF, LIMPII, OPG, IL-22, Galectin-3, MOG, Trypsin 3, SIRP α, Syndecan-4, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, Meprin A, IL-1 F10, bIG-H3, GPR115, TGFb1, Ephrin-A4, CD109, Serpin A4 Fl), IGFBP-6, HS3ST4, Aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, Cathepsin B, IGFBP-2, Semaphorin 6C, IGF-2, PDGF Rα, Sortilin, Serpin B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, Thrombomomodulin, Endoglycan, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periodin, Furin, TIMP-1, PAPP-A, Decorin, PCK1, Arylsulfatase A A) CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Lumican, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinesin, ENPP-2, CD97, CTACK, Integrin alpha 1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin 1, IGF-2R, ADAMTSL-1, Erythropoietin, Plexin D1, DNMT3ABCL-2, CL-P1, Ephrin-B3, FABP6, CHI3L1, FCRLS, TFF3, Artemin, DPPII, cIAP-1, PDGF Rb, Pentraxin 3, Angiotensinogen, Follistatin, CF VII, Persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Visfatin, Progranulin, PCSK2, GKN1, IL-18, Neprilysin, Stabilin-2, IL-17 RD, Albumin, Follistatin-like protein 1 1) MMP-10, FKBP51, LRRC4, Pref-1, Galectin-1, Troponin C, UNC5H3, FLRT2, CD314, Semaphorin 6B, Netrin-4, CD27 ligand, IL-20 Rβ, Semaphorin 6A, TSK, Cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, Norrin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF-α, PTP1B, htPAPP-A, IDO, PDGF-CC, Galanin, Activin A A) TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Glypican 2, MSP R, DSCAM, Matriptase, KIR2DL3, CD30, Siglec-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nidogen-1, CD2, Kallikrein 1, TSLP RLAMP1, TROY, VCAM-1, Siglec-11, S100A1, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R β, Thrombospondin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granzyme B, Adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Adipsin, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 α, OX40 ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Plexin A4, Semaphorin 4D, ROBO2, PDX-1, APRIL, Neuroturin, Kremen-2 (a transmembrane protein with a ring structure), EMMPRIN, ActivinRIB, Neuroligin 2, Epiregulin, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Matrilin-2, sFRP-3, p53, EphB3, NCK1, Semaphorin 7A 7A), NKp80, prolactin, cysteine ​​protease protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, semaphorin 6D, FGF-3, GATA-4, VAP-A, CHST2, pregnancy-associated plasma protein-2 (Pappalysin-2), syndecan-3, jagged 1, AKR1C4, olfactomedin-2, osteoadherin, NKp44, thyroglobulin, IL-21R,Chemerin, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin alpha 5, VAMP-2, FLRG, Ret Midkine, CD73, TRACP, proGRP, Granzyme H H), PRX2, p2'7, Siglec-6, Dendritic Cell-Associated C-type Lectin-1, CD51, Notch-1, Calreticulin, DR3, DCTN1, CDC25B, Osteoactivin, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, Cathepsin V, Testican 2, Glypican 5 5) CD6, Siglec-2, Legumin, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1β, TAFA1, Podocalyxin, RalA, CRELD2, GRAP2, SP-D, BID, GFR α-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, Catepsin L, 6-Ckine, MIS RII, Kallikrein 5, TGM3, FCAR, Contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, Biglycan, Neudesin, ILT4, uPAR, Axl, WIF-1, IL-7, Rα, GPR56, CEACAM-3, MCEMP1, FABP2, Plexin B3, MEPE, Activin RIIA, ANG-2, Cochlin, Presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, Nectin-1, GUSB, Nidogen-2, IL-17F, SR-AITAFA2, N-Cadherin, IL-17B, IL-17RC, MIP-3b, Cystatin C, Cystatin D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Enteropeptidase, Catepsin E, TSLP, TCN2, GDF-15, Epimorphin, GRKS, PD-1, Serpin A4, ADAM23, NOV, Galectin-2, Neuron surface protein 3β, TLR3, Sirtuin 2. Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphotactin, Cystatin SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, Kallikrein 14, Matrilin-3, Olig2, Kallikrein 12, CA13, IL-9, Nectin-3, MPIF-1, Cystatin S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-Selectin, Hepsin, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Semaphorin 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-1ra, Podoplanin, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenascin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2Siglec-9, ICAM-3, Cystatin SA, Galectin-4, Pepsinogen II, Desmoglein-3, Nectin-4, SCF, Serpin A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galectin-9, vWF-A2, TACE, Activin RIM, Catepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, Glypican 1, LAP (TGFb1), Gash, Contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, Nestin, TPO, ErbB3, Kirrel3, FLRT1, Galectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 Rα1, AMIGO2, HCC-1, TFPI-2, ULBP-2, Desmoglein 2, Aggrecan, Syntaxin 4 4) VAMP-1, Nectin-2, FGF-21, Flt-3, GFAP, TIM-1, Inhibin A, Cadherin-4, PIGF-2, Neurogranin, HE4, IL-23R, Galectin-7, GALNT3,GITR L, CD14, R-Spondin 2, CK19, Cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, Siglec-7, CD155, VEGF-C, TNF-α, PGRP-S, SDF-1α, PDGF-AB, GPVI, CD40, SCF-R, Thrombospondin-5, IL-1 RII, Neuropilin-2, Cadherin-13, E-Selectin, GITR, WISP-1, Renin, AgRP, MDL-1, ROBO3, RANTES, Endocan, Granulin, hCGb, Mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, Trypsin 3, ARSB, LIF Rα, BAFF R, CD157, Granzyme A, 2B4, ESAM, IL-1R4, CXCL14, IL-31, SIRP α, Uromodulin, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3, IL-32 α, TGFb3, FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Ficolin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Syntaxin 6, GRO, Bcl-w, Lipocalin-2, PDGF-AA, IL-2 Ra, Angiogenin, LYVE-1, CD4, RAGE, CDNF, Brevican, NAP-2, PU.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, procalcitonin, SOX15, kallikrein 11, BCMA, dendritic cell-associated C-lectin-2, EpCAM, HCC-4TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Syndecan-4, IL-2 Rg, MICA, Dopa Decarboxylase, NPDC-1, MCP-4, EG-VEGF, Glycoprotein V, Semaphorin 4G, IL-12p40, Total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, Endoglin, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, Galectin-8, CA4, Cystatin EM), FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7, or IL-13. In some embodiments, the therapeutic MSC secretome composition comprises one or more of the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p. p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132 -3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p,hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-mi R-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR -29a-3p、hsa-miR-29c-3p、hsa-miR-30a-5p、hsa-miR-30a-5p、hsa-miR-3 0b-5p、hsa-miR-30c-5p、hsa-mir-30d、hsa-miR-30d-5p、hsa-mir-30e、hs a-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-37 6a-3p, hsa-miR-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hs a-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p or hsa-miR-99b-5p.

[0016] One aspect of this disclosure is a method for treating graft-versus-host disease in a subject, the method comprising administering to the subject a composition comprising a therapeutic MSC secretome composition containing extracellular vesicles, wherein at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63+CD9-CD81-. In some embodiments, the therapeutic MSC secretome composition further comprises one or more of the following proteins: ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1, OPN, PAI-1, DAPP1, cathepsin B, brain signaling protein 6C, PDGF Rα, sorting protein, serine protease inhibitor B6, Dkk-3, coagulation regulatory protein, PF4, MIF, periosteal protein, furin, TIMP-1, tectonic proteoglycan, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, photoglucan, TIMP-2, SLITRK5, FAP, leptospirin, DPPII, cIAP-1, pentameric protein 3, lactone, enkephalin, albumin, galactolectin-1, UNC5H3, IL-20 Rβ, SREC-II, JAM-C, TNF. RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, convolutional protein A4, EMMPRIN, p53, brain signaling protein 7A, NKp80, cysteine ​​protease protein B, osteoadhesion, mid-term factor, calreticulin, osteoactivin, asparagine endopeptidase, TAZ, cathepsin L, RBP4, serine protease inhibitor A4, JAM-A, MCSF, LIMPII, OPG, IL-22, galactolectin-3, MOG, trypsin 3, SIRP α, cohesin-4, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, transmembrane peptidase A, IL-1 F10, bIG-H3, GPR115, TGFb1, liver glycoside-A4, CD109, serine protease inhibitor F1, IGFBP-6, HS3ST4, aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, cathepsin B, IGFBP-2, brain signaling protein 6C, IGF-2, PDGF Rα, sorting protein, serine protease inhibitor B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, coagulation regulatory protein, endosaccharide, IGFBP-3, RGM-C, PF4, MIF, TGM4, periostealin, furin, TIMP-1, PAPP-A, dermal proteoglycan, PCK1, arylsulfatase A, CD99, CA2, PRDX4, transferrin, DcR3, GP73.LAIR2, ULBP-4, photoglucan, TIMP-2, TFPI, SOX2, SLITRK5, FAP, spinal cord protein, ENPP-2, CD97, CTACK, integrin α1, EXTL3, IL-18 BPa, PD-L2, PSMA, IL-20 Ra, glyoxalase II, trypsin I, IGF-2R, ADAMTSL-1, erythropoietin, globulin D1, DNMT3A, BCL-2, CL-P1, liver glycoside-B3, FABP6, CHI3L1, FCRLS, TFF3, leptospirin, DPPII, cIAP-1, PDGF Rb, pentamin 3, angiotensinogen, follicle-stimulating hormone, CFVII, persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, endothelin, granulosa protein precursor, PCSK2, GKN1, IL-18, enkephalin, stabilizer-2, IL-17 RD, albumin, follicle-stimulating protein 1, MMP-10, FKBP51, LRRC4, Pref-1, galactoglobin-1, troponin C, UNC5H3, FLRT2, CD314, brain signaling protein 6B, cytokeratin-4, CD27 ligand, IL-20 Rβ, brain signaling protein 6A, TSK, cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, norin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Glycopropyl peptide, Activator Protein A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Phosphatidylinositol Proteoglycan 2, MSP R, DSCAM, Membrane-type Serine Protease, KIR2DL3, CD30, Salivary Agglutinin-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nestin-1, CD2, Kallikrein 1, TSLPR, LAMP1, TROY, VCAM-1, Salivary agglutinin-11, S100A1, PAR1, Thyroid peroxidase, Aminopeptidase P2, IL-1RI, ADAMS, OSM Rβ, Platelet-Reactive Protein-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granulase B, Lipocontin, IL-22BP, TPST2PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, lipoprotein, AIF, SorCS2, SULT2A1, CD39L2, insulin R, HIF-1 α, OX40 ligand, Pax3, UCH-L3, cMASP3, Langerin, desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, conglomerate A4, brain signaling protein 4D, ROBO2, PDX-1, APRIL, neuronal rankin, transmembrane protein-2 with ring structure, EMMPRIN, activator protein RIB, neuroligand 2, epithelial regulatory protein, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, extracellular matrix protein-2, sFRP-3, p53, EphB3, NCK1, brain signaling protein 7A, NKp80, prolactin, cysteine ​​agonist protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, brain signaling protein 6D, FGF-3, GATA-4, VAP-A, CHST2, pregnancy-associated plasma protein-2, cohesin-3, serrated protein 1, AKR1C4, olfactory mesrin-2, osteoadhesion, NKp44, thyroglobulin, IL-21R, chemokines, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, integrin α 5. VAMP-2, FLRG, Ret metaphase factor, CD73, TRAP, proGRP, granzyme H, PRX2, p2'7, salivary lectin-6, dendritic cell-associated C-type lectin-1, CD51, Notch-1, calreticulin, DR3, DCTN1, CDC25B, bone activator, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, cathepsin V, testosterone-2, phosphatidylinositol proteoglycan 5, CD6, salivary lectin-2, asparagine endopeptidase, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 β, TAFA1, podocyte protein, RalA, CRELD2, GRAP2, SP-D, BID, GFR α-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, cathepsin L, 6-Ckine, MIS RII, kallikrein 5, TGM3, FCAR, contactin-2, CD83, IL-1R3, SALM4, GBA3ROBO4, OSCAR, VEGF, IGS F3, disaccharide proteoglycans, neurodevelopmental factor, ILT4, uPAR, Axl, WIF-1, IL-7 Rα, GPR56, CEACAM-3, MCEMP1, FABP2, plexin B3, MEPE, activator RIIA, ANG-2, Cochlin, presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, stalkin-1, GUSB, nestin-2, IL-17F, SR-AI, TAFA2, N-cadherin, IL-17B, IL-17RC, MIP-3b, cysteine ​​protease C, cysteine ​​protease D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, intestinal peptidase, cathepsin E, TSLP, TCN2, GDF-15, epidermal morphogenetics, GRKS, PD-1, serine protease inhibitor A4, ADAM23, NOV, galactoglobulin-2, neuronal surface protein 3β, TLR3, Sirtuin2, Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, lymphocyte chemokine, cysteine ​​protease protein SN, JAM-A, calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, kallikrein 14, extracellular matrix protein-3, Oligo2, kallikrein 12, CA13, IL-9, stalk protein-3, MPIF-1, cysteine ​​protease protein S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, transmembrane serine protease, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, brain signaling protein 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-1ra, Flatfoot protein, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenosynovin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Salivary lectin-9, ICAM-3, Cysteine ​​protease inhibitor SA, Galactohemagglutinin-4, Pepsinogen II, Desmosome core protein-3, Stalk protein-4, SCF, Serine protease inhibitor A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTALEGF R, TAFAS, galactoglobulin-9, vWF-A2, TACE, activator protein RIM, cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, phosphatidylinositol proteoglycan 1, LAP (TGFb1), Gash, contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, neural epithelial stem cell protein, TPO, ErbB3, Kirrel3, FLRT1, galactoglobulin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 Rα 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, desmosome core protein 2, agglutinin, synaptic fusion protein 4, VAMP-1, stalkin-2, FGF-21, Flt-3, GFAP, TIM-1, inhibin A, cadherin-4, PIGF-2, neurogranulin, HE4, IL-23 R, galactoglobulin-7, GALNT3, GITR L, CD14, R-vertebral protein 2, CK19, cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, salivary immunoglobulin-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-1a, PDGF-AB, GPVI, CD40, SCF R, platelet-reactive protein-5, IL-1 RII, neurofeedin-2, cadherin-13, E-selectin, GITR, WISP-1, renin, AgRP, MDL-1, ROBO3, RANTES, endothelial cell-specific molecules, granzyme, hCGb, mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, trypsin 3, ARSB, LIF Rα, BAFFR, CD157, granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP α, urinary modulatory proteins, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3IL-32α, TGFb3, FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, fibrinogen-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, insulin, synaptic fusion protein 6, GRO, Bcl-w, lipocalin-2, PDGF-AA, IL-2 Ra, angiopoietin, LYVE-1, CD4, RAGE, CDNF, short proteoglycans, NAP-2, PU.1, EDAR, ADAMTS13, kynurenase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, procalcitonin, SOX15, kallikrein 11, BCMA, dendritic cell-associated C-lectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, resistin, HVEM, ENPP-7, cohesin-4, IL-2 Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, brain signaling protein 4G, IL-12p40, total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, endothelial glycoprotein, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, galactolectin-8, CA4, cysteine ​​protease protein EM, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7 or IL-13. In some implementations, the extracellular vesicles comprise one or more of the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, h sa-mir-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140,hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p、hsa-miR-148a-3p、hsa-miR-152-3p、hsa-miR-15a-5p、hsa-miR-1 5b-5p、hsa-mir-16-1、hsa-mir-16-2、hsa-miR-16-5p、hsa-miR-1'7-5 p、hsa-miR-181a-5p、hsa-miR-191-5p、hsa-miR-193a-5p、hsa-miR-19 3b-3p、hsa-miR-19'7-3p、hsa-miR-199a-3p、hsa-miR-199a-5p、hsa-m iR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p hsa-mir-24-1 hsa-mir-24-2 hsa-miR-24-3p hsa-mir-25 hsa-miR-25-3p hsa-miR-26a-5p hsa-miR-27a-3p hsa-mir-27b hsa-miR-27b-3p hsa-miR-29a-3p hsa-miR-29c-3p hsa-miR-30a-5p hsa-miR-30a-5p hsa-miR-30b-5p hsa-miR-30c-5p hsa-mir-30d hsa-miR-30 d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3p, hsa-miR-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, or hsa-miR-99b-5p. In some embodiments, the subject has received an organ transplant or allogeneic graft. In some embodiments, the subject has received a modified multi-organ allogeneic graft, a single intestinal allogeneic graft, or a single liver allogeneic graft. In some implementations, the subject exhibits one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection. In some implementations, the subject exhibits one or more signs or symptoms selected from the following: crypt cell apoptosis, crypt loss, ulceration, allogeneic graft cell rejection, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof. In some implementations, the subject is receiving corticosteroids. In some implementations, the subject is receiving high-dose corticosteroids. In some implementations, subjects experience improvement in one or more of the following signs or symptoms following treatment: crypt cell apoptosis, crypt loss, ulcer formation, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, or combinations thereof. In some implementations, subjects show improvement within 24 hours of administration of the treatment product.

[0017] One aspect of this disclosure is a method for treating graft-versus-host disease in a subject, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles (EVs), wherein the therapeutic MSC secretome comprises: (i) ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1, OPN, PAI-1, DAPP1, cathepsin B, brain signaling protein 6C, PDGF R α, Sorting protein, Serine protease inhibitor B6, Dkk-3, Coagulation regulator protein, PF4, MIF, Periostrin, Flin protease, TIMP-1, Gynostemma pentaglycoside, PCK1, CD99, CD63, CD9, CD81, Transferrin, DcR3, Optical proteoglycan, TIMP-2, SLITRK5, FAP, Neurosphingomyelin, DPPII, cIAP-1, Pentanoic acid 3, Lactopeptidase, Enkephalin, Albumin, Galactoglobin-1, UNC5H3, IL-20 Rβ, SREC-II, JAM-C, TNF-α, htPAPP-A, eNOS, MSP-R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, convolutional protein A4, EMMPRIN, p53, brain signaling protein 7A, NKp80, cysteine ​​protease protein B, osteoadhesion, mid-term factor, calreticulin, osteoactivin, asparagine endopeptidase, TAZ, cathepsin L, RBP4, serine protease inhibitor A4, JAM-A, MCSF, LIMPII, OPG, IL-22, galactoglobulin-3, MOG, trypsin 3, SIRP α, cohesin-4, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, transmembrane peptidase A, IL-1 F10, bIG-H3, GPR115, TGFb1, liver glycoside A4, CD109, serine protease inhibitor F1, IGFBP-6, HS3ST4, aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, cathepsin B, IGFBP-2, brain signaling protein 6C, IGF-2, PDGFR α, sorting protein, serine protease inhibitor B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, coagulation regulatory protein, endosaccharides, IGFBP-3, RGM-C, PF4, MIF, TGM4, periosteal protein, furin protease, TIMP-1, PAPP-A, dermal proteoglycans, PCK1, arylsulfatase A, CD99, CA2, PRDX4, transferrin, DcR3, GP73, LAIR2, ULBP-4, optical proteoglycans, TIMP-2, TFPI, SOX2, SLITRK5, FAP, spinal proteinENPP-2, CD97, CTACK, integrin α1, EXTL3, IL-18, BPa, PD-L2, PSMA, IL-20 Ra, glyoxalase II, trypsin I, IGF-2R, ADAMTSL-1, erythropoietin, plexin D1, DNMT3A, BCL-2, CL-P1, hepatocyte glycoside-B3, FABP6, CHI3L1, FCRLS, TFF3, leptospirin, DPPII, cIAP-1, PDGF Rb, pentameric protein 3, angiotensinogen, follicle-stimulating hormone, CF VII, persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, endothelin, granulosin precursor, PCSK2, GKN1, IL-18, enkephalin, stabilizer-2, IL-17 RD, albumin, follicle-stimulating protein 1, MMP-10, FKBP51, LRRC4, Pref-1, galactoglobin-1, troponin C, UNC5H3, FLRT2, CD314, brain signaling protein 6B, cytokinin-4, CD27 ligand, IL-20 Rβ, brain signaling protein 6A, TSK, cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, norin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Glycopropyl peptide, Activator Protein A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Phosphatidylinositol Proteoglycan 2, MSP R, DSCAM, Membrane-type Serine Protease, KIR2DL3, CD30, Salivary Agglutinin-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nestin-1, CD2, Kallikrein 1, TSLPR, LAMP1, TROY, VCAM-1, Salivary agglutinin-11, S100A1, PAR1, Thyroid peroxidase, Aminopeptidase P2, IL-1RI, ADAMS, OSM Rβ, Platelet-Reactive Protein-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granulase B, Lipocontin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, LipoproteinAIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 α, OX40 ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Convolutional protein A4, Brain signaling protein 4D, ROBO2, PDX-1, APRIL, Neural rank protein, cyclic transmembrane protein-2, EMMPRIN, activator protein RIB, neuroligand 2, epithelial regulatory protein, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Extracellular matrix protein-2, sFRP-3, p53, EphB3, NCK1, Brain signaling protein 7A, NKp80, Prolactin, Cysteine ​​protease protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Brain signaling protein 6D, FGF-3, GATA-4, VAP-A, CHST2, Pregnancy-associated plasma protein-2, Cohesin-3, Serpentin-1, AKR1C4, Oligosin-2, Osteoadhesin, NKp44, Thyroglobulin, IL-21R, Chemokines, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin α 5. VAMP-2, FLRG, Ret metaphase factor, CD73, TRAP, proGRP, granzyme H, PRX2, p2'7, salivary lectin-6, dendritic cell-associated C-type lectin-1, CD51, Notch-1, calreticulin, DR3, DCTN1, CDC25B, bone activator, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, cathepsin V, testosterone-2, phosphatidylinositol proteoglycan 5, CD6, salivary lectin-2, asparagine endopeptidase, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 β, TAFA1, podocyte protein, RalA, CRELD2, GRAP2, SP-D, BID, GFR α-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, cathepsin L, 6-Ckine, MIS RII, kallikrein 5, TGM3, FCAR, contactin-2, CD83, IL-1R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, disaccharide proteoglycan, neurodevelopmental factor, ILT4, uPAR, Axl, WIF-1IL-7 Rα, GPR56, CEACAM-3, MCEMP1, FABP2, plexin B3, MEPE, activator RIIA, ANG-2, Cochlin, progerin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, stalkin-1, GUSB, nestin-2, IL-17F, SR-AI, TAFA2, N-cadherin, IL-17B, IL-17RC, MIP-3b, cysteine ​​protease C, cysteine ​​protease D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, intestinal peptidase, cathepsin E, TSLP, TCN2, GDF-15, epidermal morphogenetics, GRKS, PD-1, serine protease inhibitor A4, ADAM23, NOV, galactoglobulin-2, neuronal surface protein 3β, TLR3, Sirtuin2, Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, lymphocyte chemokine, cysteine ​​protease protein SN, JAM-A, calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, kallikrein 14, extracellular matrix protein-3, Oligo2, kallikrein 12, CA13, IL-9, stalk protein-3, MPIF-1, cysteine ​​protease protein S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, transmembrane serine protease, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, brain signaling protein 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-1ra, Flatfoot protein, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenosynovin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Salivary lectin-9, ICAM-3, Cysteine ​​protease inhibitor SA, Galactohemagglutinin-4, Pepsinogen II, Desmosome core protein-3, Stalk protein-4, SCF, Serine protease inhibitor A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galactohemagglutinin-9, vWF-A2, TACE, Activator protein RIM, Cathepsin S, LDL RBMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, phosphatidylinositol proteoglycan 1, LAP (TGFb1), Gash, contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, neural epithelial stem cell protein, TPO, ErbB3, Kirrel3, FLRT1, galactoglobulin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 Rα 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, desmosome core protein 2, agglutinin, synaptic fusion protein 4, VAMP-1, stalkin-2, FGF-21, Flt-3, GFAP, TIM-1, inhibin A, cadherin-4, PIGF-2, neurogranulin, HE4, IL-23 R, galactoglobulin-7, GALNT3, GITR L, CD14, R-vertebral protein 2, CK19, cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, salivary immunoglobulin-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-1a, PDGF-AB, GPVI, CD40, SCF R, platelet-reactive protein-5, IL-1 RII, neurofeedin-2, cadherin-13, E-selectin, GITR, WISP-1, renin, AgRP, MDL-1, ROBO3, RANTES, endothelial cell-specific molecules, granzyme, hCGb, mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, trypsin 3, ARSB, LIF Rα, BAFFR, CD157, granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP α, urinary modulatory proteins, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3, IL-32 α, TGFb3, FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, fibrinogen-1, Reg4ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Synaptic Fusion Protein 6, GRO, Bcl-w, Lipocrine-2, PDGF-AA, IL-2 Ra, Angiopoietin, LYVE-1, CD4, RAGE, CDNF, Short Proteoglycan, NAP-2, PU.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dendritic Cell-Associated C-Lectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Cohesin-Glycan-4, IL-2 Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, brain signaling protein 4G, IL-12p40, total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, endothelial glycoprotein, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, galactolectin-8, CA4, cysteine ​​protease protein EM, FUT8, B7-H 3. GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7 or IL-13 or a combination of two or more thereof; (ii) hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hs a-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir- 10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-mi R-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p , hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p,hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-mi R-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa- mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p hsa-miR-320a hsa-miR-342-3p hsa-miR-345-5p hsa-miR-34a-5p hsa-miR-361-5p hsa-miR-376a-3p hsa-miR-376c-3p hsa-miR-423-3p hsa -miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663ahsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p, or hsa-miR-99b-5p, or a combination of two or more thereof; or both (iii), (i), and (ii). Another aspect of this disclosure is a method for treating graft-versus-host disease, comprising administering a therapeutic MSC secretome composition to a subject. Another aspect of this disclosure is a method for treating allogeneic graft rejection, comprising administering a therapeutic MSC secretome composition to a subject. Another aspect of this disclosure is a method for alleviating symptoms associated with graft-versus-host disease in a subject, comprising administering a therapeutic MSC secretome composition to the subject. Another aspect of this disclosure is a method for improving signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection selected from one or more of the following: crypt cell apoptosis, crypt loss, ulceration, allogeneic graft cell rejection, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, pruritus, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof. In some embodiments, the extracellular vesicles in the therapeutic MSC secretome composition are CD63+CD9-CD81-. In some implementations, the therapeutic MSC secretion group includes: (i) ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1, OPN, PAI-1, DAPP1, cathepsin B, brain signaling protein 6C, PDGF Rα, sorting protein, serine protease inhibitor B6, Dkk-3, coagulation regulatory protein, PF4, MIF, periostealin, furin, TIMP-1, tretinoin, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, photoglucan, TIMP-2, SLITRK5, FAP, leptospirin, DPPII, cIAP-1, pentamericin 3, lactone, enkephalin, albumin, galactolectin-1, UNC5H3, IL-20 Rβ, SREC-II, JAM-C, TNFRI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, pyrine A4, EMMPRIN, p53, brain signaling protein 7A, NKp80, cysteine ​​protease protein B, osteoadhesion, mid-term factor, calreticulin, osteoactivin, asparagine endopeptidase, TAZ, cathepsin L,RBP4, serine protease inhibitor A4, JAM-A, MCSF, LIMPII, OPG, IL-22, galactoglobulin-3, MOG, trypsin 3, SIRP α, cohesin-4, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, transmembrane peptidase A, IL-1 F10, big-H3, GPR115, TGFb1, liver glycoside A4, CD109, serine protease inhibitor F1, IGFBP-6, HS3ST4, aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, cathepsin B, IGFBP-2, brain signaling protein 6C, IGF-2, PDGF R α, sorting protein, serine protease inhibitor B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, coagulation regulatory protein, endosaccharides, IGFBP-3, RGM-C, PF4, MIF, TGM4, periosteal protein, furin, TIMP-1, PAPP-A, dermal proteoglycan, PCK1, arylsulfatase A, CD99, CA2, PRDX4, transferrin, DCR3, GP73, LAIR2, ULBP-4, optical proteoglycan, TIMP-2, TFPI, SOX2, SLITRK5, FAP, spinal cord protein, ENPP-2, CD97, CTACK, integrin α1, EXTL3, IL-18, BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin I, IGF-2R, ADAMTS L1-1, Erythropoietin, Convolutional D1, DNMT3A, BCL-2, CL-P1, Hepatocyte-B3, FABP6, CHI3L1, FCRLS, TFF3, Nephlebotomycin, DPPII, cIAP-1, PDGF Rb, Pentanoic Acid 3, Angiotensinogen, Follicle-Stapressin, CF VII, Persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Lactone, Granulosin Progenitor, PCSK2, GKN1, IL-18, Enkephalin, Stabilin-2, IL-17 RD, albumin, follicle-stimulating protein 1, MMP-10, FKBP51, LRRC4, Pref-1, galactoglobin-1, troponin C, UNC5H3, FLRT2, CD314, brain signaling protein 6B, cytokinin-4, CD27 ligand, IL-20 Rβ, brain signaling protein 6A, TSK, cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, norin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF-α, PTP1B.htPAPP-A, IDO, PDGF-CC, Glycolipide, Activator Protein A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Phosphatidylinositol Proteoglycan 2, MSP R, DSCAM, Membrane-type Serine Protease, KIR2DL3, CD30, Salivary Agglutinin-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nestin-1, CD2, Kallikrein 1, TSLP R, LAMP1, TROY, VCAM-1, Salivary Agglutinin-11, S100A1, PAR1, Thyroid Peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM Rβ, thromboretin-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), granzyme B, adiponectin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, lipoprotein, AIF, SorCS2, SULT2A1, CD39L2, insulin R, HIF-1 α, OX40 ligand, Pax3, UCH-L3, cMASP3, Langerin, desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, conglomerate A4, brain signaling protein 4D, ROBO2, PDX-1, APRIL, neuronal rank protein, transmembrane protein-2 with ring structure, EMMPRIN, activating protein RIB, neuroligand 2, epithelial regulatory protein, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, extracellular matrix protein-2, sFRP-3, p53, EphB3, NCK1, brain signaling protein 7A, NKp80, prolactin, cysteine ​​agonist protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Brain signaling protein 6D, FGF-3, GATA-4, VAP-A, CHST2, Pregnancy-associated plasma protein-2, cohesin-3, serrated protein 1, AKR1C4, olfactory mesentery protein-2, osteoadhesion, NKp44, thyroglobulin, IL-21R, chemokines, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, integrin α5, VAMP-2, FLRG, Ret metaphase factor, CD73, TRACP, proGRP, granzyme H, PRX2.p27, salivary lectin-6, dendritic cell-associated C-type lectin-1, CD51, Notch-1, calreticulin, DR3, DCTN1, CDC25B, bone activator, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, cathepsin V, testosterone-2, phosphatidylinositol proteoglycan-5, CD6, salivary lectin-2, asparagine endopeptidase, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1β, TAFA1, podocyte calyx protein, RalA, CRELD2, GRAP2, SP-D, BID, GFR α-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, cathepsin L, 6-Ckine, MIS RII, kallikrein 5, TGM3, FCAR, contactin-2, CD83, IL-1, R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, disaccharide proteoglycan, neurodevelopmental factor, ILT4, uPAR, Axl, WIF-1, IL-7, Rα, GPR56, CEACAM-3, MCEMP1, FABP2, plexin B3, MEPE, activator RIIA, ANG-2, Cochlin, progerin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, stalkin-1, GUSB, nestin-2, IL-17F, SR-AI, TAFA2, N-cadherin, IL-17B, IL-17 RC, MIP-3b, Cysteine ​​protease protein C, Cysteine ​​protease protein D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, Prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, Intestinal peptidase, Cathepsin E, TSLP, TCN2, GDF-15, Epidermal morphogenetics, GRKS, PD-1, Serine protease inhibitor A4, ADAM23, NOV, Galactoglobin-2, Neuronal surface protein 3β, TLR3, Sirtuin 2, Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, Lymphocyte chemokine, Cysteine ​​protease protein SN, JAM-A, Calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, kallikrein 14, extracellular matrix protein-3, Olig2, kallikrein 12, CA13, IL-9, stalk protein-3MPIF-1, Cysteine ​​protease S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, transmembrane serine protease, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Brain signaling protein 4C, SorCS3, Tie-1, IL-31RA, Arginase 1, POGLUT1, IL-1ra, Flatfoot protein, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenosynovin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Salivary lectin-9, ICAM-3, Cysteine ​​protease inhibitor SA, Galactohemagglutinin-4, Pepsinogen II, Desmosome core protein-3, Stalk protein-4, SCF, Serine protease inhibitor A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, galactoglobulin-9, vWF-A2, TACE, activator protein RIM, cathepsin S, LDLR, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, phosphatidylinositol proteoglycan 1, LAP (TGFb1), Gash, contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, neural epithelial stem cell protein, TPO, ErbB3, Kirrel3, FLRT1, galactolectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGFR2, Tie-2, IL-15R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22Rα1, AMIGO2, HCC-1, TFPI-2, ULBP-2, desmosome core protein 2, agglutinin, synaptic fusion protein 4, VAMP-1, stalkin-2, FGF-21, Flt-3, GFAP, TIM-1, inhibin A, cadherin-4, P1GF-2, neurogranulin, HE4.IL-23R, galactoglobulin-7, GALNT3, GITR L, CD14, R-vertebral protein 2, CK19, cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, salivary immunoglobulin-7, CD155, VEGF-C, TNF-α, PGRP-S, SDF-1a, PDGF-AB, GPVI, CD40, SCFR, thromboretin-5, IL-1 α, neurolin-2, cadherin-13, E-selectin, GITR, WISP-1, renin, AgRP, MDL-1, ROBO3, RANTES, endothelial cell-specific molecules, granulin, hCGb, mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, trypsin 3, ARSB, LIF Rα, BAFF R, CD157, Granulase A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP α, Urinary Modulatory Protein, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3, IL-32 α, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, Fibrin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Synaptic Fusion Protein 6, GRO, Bcl-w, Lipocrine-2, PDGF-AA, IL-2 Ra, Angiopoietin, LYVE-1, CD4, RAGE, CDNF, Short Proteoglycan, NAP-2, PU.1, EDAR, ADAMTS13, Kynurenase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, procalcitonin, SOX15, kallikrein 11, BCMA, dendritic cell-associated C-lectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, resistin, HVEM, ENPP-7, cohesin-4, IL-2 Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, brain signaling protein 4G, IL-12p40, total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, endothelial glycoprotein, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, galactolectin-8, CA4, cysteine ​​protease protein EM, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13Kallikrein 7 or IL-13 or a combination of two or more thereof; (ii) hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1’7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19’7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p,hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR- 29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hs a-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3p, hsa-m iR-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486- 1. hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR -874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p or hsa-miR-99b-5p, or a combination of two or more thereof; or (iii) both (i) and (ii). In some embodiments, the therapeutic MSC secretome composition is prepared by a method comprising the following steps: (a) culturing bone marrow-derived MSCs to produce MSC conditioned medium under conditions of (i) oxygen tension below 5%; and (ii) a medium with a pH below 7; (b) harvesting the MSC conditioned medium; and (c) formulating the MSC conditioned medium to produce the therapeutic MSC secretome composition, wherein the therapeutic MSC secretome composition comprises proteins and extracellular vesicles produced by bone marrow-derived MSCs in step (a). In some embodiments, the subject has received an organ transplant or allogeneic graft. In some embodiments, the subject has received a modified multi-organ allogeneic graft, a single intestinal allogeneic graft, or a single liver allogeneic graft. In some embodiments, the subject exhibits one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection. In some embodiments, the subject exhibits one or more signs or symptoms selected from: crypt cell apoptosis, crypt loss, ulceration,Cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, blisters, peeling, tenderness, liver failure, ulceration, and combinations thereof. In some embodiments, the composition comprises 15 mL of a therapeutic MSC secretory group composition. In some embodiments, the composition is administered by intravenous injection.

[0018] The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution may be about 100 mL. The number of EVs in the composition may be about 10 million to about 1 trillion. The number of EVs in the composition may be about 1 trillion. The number of EVs in the composition may be about 10 billion to about 1 trillion.

[0019] One aspect of this disclosure is a method for treating graft-versus-host disease in a subject, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretory composition containing extracellular vesicles (EVs), wherein the composition comprises 15 mL of the therapeutic MSC secretory composition. Another aspect of this disclosure is a method for treating graft-versus-host disease in a subject, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretory composition containing extracellular vesicles (EVs), wherein the administration improves one or more signs or symptoms selected from: crypt cell apoptosis, crypt loss, ulceration, allogeneic graft cell rejection, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof. In some embodiments, the administration improves inflammation.

[0020] One aspect of this disclosure is the use of a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles in graft-versus-host disease of a therapeutic subject, wherein at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63+CD9-CD81-.

[0021] One aspect of this disclosure is the use of a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles (EVs) for the treatment of graft-versus-host disease in subjects of need, wherein the therapeutic mesenchymal stem cell (MSC) secretome composition comprises: (i) ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1, OPN, PAI-1, DAPP1, cathepsin B, brain signaling protein 6C, PDGF R α, Sorting protein, Serine protease inhibitor B6, Dkk-3, Coagulation regulator protein, PF4, MIF, Periostrin, Flin protease, TIMP-1, Gynostemma pentaglycoside, PCK1, CD99, CD63, CD9, CD81, Transferrin, DcR3, Optical proteoglycan, TIMP-2, SLITRK5, FAP, Neurosphingomyelin, DPPII, cIAP-1, Pentanoic acid 3, Lactopeptidase, Enkephalin, Albumin, Galactoglobin-1, UNC5H3, IL-20 Rβ, SREC-II, JAM-C, TNF-α, htPAPP-A, eNOS, MSP-R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, convolutional protein A4, EMMPRIN, p53, brain signaling protein 7A, NKp80, cysteine ​​protease protein B, osteoadhesion, mid-term factor, calreticulin, osteoactivin, asparagine endopeptidase, TAZ, cathepsin L, RBP4, serine protease inhibitor A4, JAM-A, MCSF, LIMPII, OPG, IL-22, galactoglobulin-3, MOG, trypsin 3, SIRP α, cohesin-4, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, transmembrane peptidase A, IL-1 F10, bIG-H3, GPR115, TGFb1, liver glycoside A4, CD109, serine protease inhibitor F1, IGFBP-6, HS3ST4, aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, cathepsin B, IGFBP-2, brain signaling protein 6C, IGF-2, PDGFR α, sorting protein, serine protease inhibitor B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, coagulation regulatory protein, endosaccharides, IGFBP-3, RGM-C, PF4, MIF, TGM4, periosteal protein, furin protease, TIMP-1, PAPP-A, dermal proteoglycans, PCK1, arylsulfatase A, CD99, CA2, PRDX4, transferrin, DcR3, GP73, LAIR2, ULBP-4, optical proteoglycans, TIMP-2, TFPI, SOX2, SLITRK5, FAP, spinal proteinENPP-2, CD97, CTACK, integrin α1, EXTL3, IL-18, BPa, PD-L2, PSMA, IL-20 Ra, glyoxalase II, trypsin I, IGF-2R, ADAMTSL-1, erythropoietin, plexin D1, DNMT3A, BCL-2, CL-P1, hepatocyte glycoside-B3, FABP6, CHI3L1, FCRLS, TFF3, leptospirin, DPPII, cIAP-1, PDGF Rb, pentameric protein 3, angiotensinogen, follicle-stimulating hormone, CF VII, persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, endothelin, granulosin precursor, PCSK2, GKN1, IL-18, enkephalin, stabilizer-2, IL-17 RD, albumin, follicle-stimulating protein 1, MMP-10, FKBP51, LRRC4, Pref-1, galactoglobin-1, troponin C, UNC5H3, FLRT2, CD314, brain signaling protein 6B, cytokinin-4, CD27 ligand, IL-20 Rβ, brain signaling protein 6A, TSK, cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, norin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Glycopropyl peptide, Activator Protein A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Phosphatidylinositol Proteoglycan 2, MSP R, DSCAM, Membrane-type Serine Protease, KIR2DL3, CD30, Salivary Agglutinin-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nestin-1, CD2, Kallikrein 1, TSLPR, LAMP1, TROY, VCAM-1, Salivary agglutinin-11, S100A1, PAR1, Thyroid peroxidase, Aminopeptidase P2, IL-1RI, ADAMS, OSM Rβ, Platelet-Reactive Protein-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granulase B, Lipocontin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, LipoproteinAIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 α, OX40 ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Convolutional protein A4, Brain signaling protein 4D, ROBO2, PDX-1, APRIL, Neural rank protein, cyclic transmembrane protein-2, EMMPRIN, activator protein RIB, neuroligand 2, epithelial regulatory protein, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Extracellular matrix protein-2, sFRP-3, p53, EphB3, NCK1, Brain signaling protein 7A, NKp80, Prolactin, Cysteine ​​protease protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Brain signaling protein 6D, FGF-3, GATA-4, VAP-A, CHST2, Pregnancy-associated plasma protein-2, Cohesin-3, Serpentin-1, AKR1C4, Oligosin-2, Osteoadhesin, NKp44, Thyroglobulin, IL-21R, Chemokines, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin α 5. VAMP-2, FLRG, Ret metaphase factor, CD73, TRAP, proGRP, granzyme H, PRX2, p2'7, salivary lectin-6, dendritic cell-associated C-type lectin-1, CD51, Notch-1, calreticulin, DR3, DCTN1, CDC25B, bone activator, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, cathepsin V, testosterone-2, phosphatidylinositol proteoglycan 5, CD6, salivary lectin-2, asparagine endopeptidase, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 β, TAFA1, podocyte protein, RalA, CRELD2, GRAP2, SP-D, BID, GFR α-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, cathepsin L, 6-Ckine, MIS RII, kallikrein 5, TGM3, FCAR, contactin-2, CD83, IL-1R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, disaccharide proteoglycan, neurodevelopmental factor, ILT4, uPAR, Axl, WIF-1IL-7 Rα, GPR56, CEACAM-3, MCEMP1, FABP2, plexin B3, MEPE, activator RIIA, ANG-2, Cochlin, progerin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, stalkin-1, GUSB, nestin-2, IL-17F, SR-AI, TAFA2, N-cadherin, IL-17B, IL-17RC, MIP-3b, cysteine ​​protease C, cysteine ​​protease D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, intestinal peptidase, cathepsin E, TSLP, TCN2, GDF-15, epidermal morphogenetics, GRKS, PD-1, serine protease inhibitor A4, ADAM23, NOV, galactoglobulin-2, neuronal surface protein 3β, TLR3, Sirtuin2, Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, lymphocyte chemokine, cysteine ​​protease protein SN, JAM-A, calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, kallikrein 14, extracellular matrix protein-3, Oligo2, kallikrein 12, CA13, IL-9, stalk protein-3, MPIF-1, cysteine ​​protease protein S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, transmembrane serine protease, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, brain signaling protein 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-1ra, Flatfoot protein, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenosynovin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Salivary lectin-9, ICAM-3, Cysteine ​​protease inhibitor SA, Galactohemagglutinin-4, Pepsinogen II, Desmosome core protein-3, Stalk protein-4, SCF, Serine protease inhibitor A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galactohemagglutinin-9, vWF-A2, TACE, Activator protein RIM, Cathepsin S, LDL RBMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, phosphatidylinositol proteoglycan 1, LAP (TGFb1), Gash, contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, neural epithelial stem cell protein, TPO, ErbB3, Kirrel3, FLRT1, galactoglobulin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 Rα 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, desmosome core protein 2, agglutinin, synaptic fusion protein 4, VAMP-1, stalkin-2, FGF-21, Flt-3, GFAP, TIM-1, inhibin A, cadherin-4, PIGF-2, neurogranulin, HE4, IL-23 R, galactoglobulin-7, GALNT3, GITR L, CD14, R-vertebral protein 2, CK19, cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, salivary immunoglobulin-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-1a, PDGF-AB, GPVI, CD40, SCF R, platelet-reactive protein-5, IL-1 RII, neurofeedin-2, cadherin-13, E-selectin, GITR, WISP-1, renin, AgRP, MDL-1, ROBO3, RANTES, endothelial cell-specific molecules, granzyme, hCGb, mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, trypsin 3, ARSB, LIF Rα, BAFFR, CD157, granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP α, urinary modulatory proteins, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3, IL-32 α, TGFb3, FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, fibrinogen-1, Reg4ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Synaptic Fusion Protein 6, GRO, Bcl-w, Lipocrine-2, PDGF-AA, IL-2 Ra, Angiopoietin, LYVE-1, CD4, RAGE, CDNF, Short Proteoglycan, NAP-2, PU.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dendritic Cell-Associated C-Lectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Cohesin-Glycan-4, IL-2 Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, brain signaling protein 4G, IL-12p40, total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, endothelial glycoprotein, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, galactolectin-8, CA4, cysteine ​​protease protein EM, FUT8, B7-H 3. GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7 or IL-13 or a combination of two or more thereof; (ii) hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hs a-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir- 10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-mi R-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p , hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p,hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-mi R-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa- mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p hsa-miR-320a hsa-miR-342-3p hsa-miR-345-5p hsa-miR-34a-5p hsa-miR-361-5p hsa-miR-376a-3p hsa-miR-376c-3p hsa-miR-423-3p hsa -miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663ahsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p or hsa-miR-99b-5p, or a combination of two or more thereof; or both (iii), (i) and (ii).

[0022] One aspect of this disclosure is the use of a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles (EVs) in the treatment of graft-versus-host disease in a subject, wherein the composition comprises 15 mL of the therapeutic MSC secretome composition. The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution may be about 100 mL. The number of EVs in the composition may be about 10 million to about 1 trillion. The number of EVs in the composition may be about 1 trillion. The number of EVs in the composition may be about 10 billion to about 1 trillion. Attached Figure Description

[0023] The novel features of this disclosure are set forth in the appended claims. A better understanding of the features and advantages of this disclosure will be obtained by referring to the following detailed description and accompanying drawings, which illustrate illustrative embodiments utilizing the principles of this disclosure, in which: Figure 1 Histological images showing improvement in allogeneic graft inflammation and relief of acute cellular rejection following administration of the therapeutic product are displayed. Detailed Implementation

[0024] Therapeutic Composition Extracellular vesicles (EVs) are membrane-bound globules containing proteins and RNA (exosomes are a subset of them). Exosomes are small (e.g., 30–170 nm) diameter lipid bilayer vesicles secreted by cells to initiate paracrine communication. Other EV populations originate directly from the plasma membrane or form during apoptosis (apoptotic bodies). This article discloses compositions containing therapeutically effective amounts of MSC secretory components (e.g., including but not limited to MSC growth factors, MSC exosomes, MSC extracts, and / or compositions containing extracellular vesicles).

[0025] The exemplary compositions described herein comprise therapeutic compositions comprising biomolecules (which may include proteins, lipids, and / or ribonucleic acid) secreted from mesenchymal lineage cells and / or extracellular vesicles containing biomolecules. In one aspect, the therapeutic composition comprises a therapeutically effective amount of a component secreted from MSCs (i.e., the MSC secretome), such as, but not limited to, MSC growth factors, MSC exosomes, MSC extracts, and / or compositions comprising extracellular vesicles. In an exemplary embodiment, the MSC is bone marrow MSC. The MSC secretome can be purified or otherwise isolated from MSC growth and / or culture conditions. For example, the MSC secretome may comprise exosomes isolated from MSC cultures. Thus, references to the MSC secretome include purified secretome components, such as exosomes or extracellular vesicles. Purified secretome components may be therapeutic compositions. Partially purified secretome components may be therapeutic compositions. For example, a therapeutic composition may comprise extracellular vesicles from the MSC secretome and one or more growth factors. The therapeutic composition may comprise reconstructed extracellular vesicles, such as those reconstructed from lyophilized or otherwise powdered or dried compositions. The therapeutic composition may contain reconstructed extracellular vesicles and one or more growth factors, for example, reconstructed from a lyophilized or otherwise powdered or dried composition.

[0026] In some embodiments, the MSC secretome comprises an MSC growth factor composition. In some embodiments, the MSC secretome is an MSC growth factor composition. The MSC growth factor composition may comprise one or more growth factors secreted from MSCs. One or more of the one or more growth factors secreted from MSCs may be present in exosomes secreted from MSCs. One or more of the one or more growth factors secreted from MSCs may be present in extracellular vesicles secreted from MSCs. One or more of the one or more growth factors secreted from MSCs may not be present in exosomes or extracellular vesicles secreted from MSCs. In an exemplary embodiment, the MSC is a bone marrow MSC.

[0027] In some embodiments, the MSC secretome comprises an MSC exosome composition. In some embodiments, the MSC secretome is an MSC exosome composition. The MSC exosome composition may comprise one or more exosomes secreted from MSCs. MSC exosomes may comprise one or more biomolecules, such as peptides, polypeptides, proteins, siRNA, shRNA, and / or microRNAs (miRNAs). MSC exosomes may comprise growth factors. Growth factors may be present in one or more exosomes secreted from MSCs. Growth factors may not be present in one or more exosomes secreted from MSCs. MSC exosomes may comprise nucleic acids, such as miRNAs. Nucleic acids may be present in one or more exosomes secreted from MSCs. Nucleic acids may not be present in one or more exosomes secreted from MSCs. In an exemplary embodiment, the MSC is a bone marrow MSC.

[0028] In some embodiments, the MSC secretome comprises an MSC extracellular vesicle composition. In some embodiments, the MSC secretome is an MSC extracellular vesicle composition. The MSC extracellular vesicle composition may comprise one or more extracellular vesicles secreted from MSCs. The MSC extracellular vesicle composition may comprise one or more biomolecules, such as peptides, polypeptides, proteins, siRNA, shRNA, and / or microRNAs (miRNAs). MSC extracellular vesicles may contain growth factors. Growth factors may be present within one or more extracellular vesicles secreted from MSCs. Growth factors may not be present within one or more extracellular vesicles secreted from MSCs. MSC extracellular vesicles may contain nucleic acids, such as miRNAs. Nucleic acids may be present within one or more extracellular vesicles secreted from MSCs. Nucleic acids may not be present within one or more extracellular vesicles secreted from MSCs. In an exemplary embodiment, the MSC is a bone marrow MSC.

[0029] In some embodiments, the MSC secretome comprises an MSC extract composition. In an exemplary embodiment, the MSC is a bone marrow MSC. In some embodiments, the MSC secretome is an MSC extract composition. The MSC extract composition may comprise one or more biomolecules, such as peptides, polypeptides, proteins, siRNA, shRNA, and / or microRNAs (miRNAs). The MSC extract composition may comprise exosomes and one or more biomolecules, wherein the exosomes are secreted from MSCs and one or more biomolecules are secreted from MSCs. One or more of the one or more biomolecules may be present in the exosomes. One or more of the one or more biomolecules may not be present in the exosomes. The MSC extract composition may comprise extracellular vesicles and one or more biomolecules, wherein the extracellular vesicles are secreted from MSCs and one or more biomolecules are secreted from MSCs. One or more of the one or more biomolecules may be present within the extracellular vesicles. One or more of the one or more biomolecules may not be present in the extracellular vesicles. One of the one or more biomolecules may be a growth factor. One of the one or more biomolecules may be a nucleic acid, such as miRNA.

[0030] In some embodiments, the MSC secretome comprises an MSC growth factor composition and an MSC exosome composition. In an exemplary embodiment, the MSC is a bone marrow MSC. The MSC growth factor composition may comprise one or more growth factors secreted from MSCs. One or more of the one or more growth factors secreted from MSCs may be present in the MSC exosome composition. One or more of the one or more growth factors secreted from MSCs may not be present in the exosomes. The MSC exosome composition may comprise one or more exosomes secreted from MSCs. MSC exosomes may comprise one or more biomolecules, such as peptides, polypeptides, proteins, siRNA, shRNA, and / or microRNAs (miRNAs). One of the one or more biomolecules may be a nucleic acid, such as miRNA.

[0031] In some embodiments, the MSC secretory group comprises an MSC growth factor composition and an MSC extracellular vesicle composition. In an exemplary embodiment, the MSC is a bone marrow MSC. The MSC growth factor composition may comprise one or more growth factors secreted from MSCs. One or more of the one or more growth factors secreted from MSCs may be present in the MSC extracellular vesicle composition. One or more of the one or more growth factors secreted from MSCs may not be present in the extracellular vesicles. The MSC extracellular vesicles may comprise one or more extracellular vesicles secreted from MSCs. The MSC extracellular vesicles may comprise one or more biomolecules, such as peptides, polypeptides, proteins, siRNA, shRNA, and / or microRNAs (miRNAs). One of the one or more biomolecules may be a nucleic acid, such as miRNA.

[0032] In some embodiments, the therapeutic composition (e.g., an MSC secretome composition) comprises one or more peptide biomolecules. At least one of the one or more peptide biomolecules may be a growth factor. For example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 or more proteins may be present in the MSC secretome. Non-restricted exemplary growth factors or growth factor-related proteins that may be present in the MSC secretome include: ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1, OPN, PAI-1, DAPP1, cathepsin B, brain signaling protein 6C, PDGF Rα, sorting protein, serine protease inhibitor B6, Dkk-3, coagulation regulator protein, PF4, MIF, periostealin, furin, TIMP-1, gypsum proteoglycan, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, photoglucan, TIMP-2, SLITRK5, FAP, leptospirin, DPPII, cIAP-1, pentamericin 3, lactone, enkephalin, albumin, galactolectin-1, UNC5H3, IL-20 Rβ, SREC-II, JAM-C, TNF. RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, conglomerate A4, EMMPRIN, p53, brain signaling protein 7A, NKp80, cysteine ​​protease protein B, osteoadhesion, mid-term factor, calreticulin, osteoactivin, asparagine endopeptidase, TAZ, cathepsin L, RBP4, serine protease inhibitor A4, JAM-A, MCSF, LIMPII, OPG, IL-22, galactoglobulin-3, MOG, trypsin 3, SIRP α, and cohesin-glycan-4. Non-restricted exemplary proteins that may be present in the MSC secretome include: ferritin, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, transmembrane peptidase A, IL-1 F10, bIG-H3, GPR115, TGFb1, hepatocyte glycoside A4, CD109, serine protease inhibitor F1, IGFBP-6, HS3ST4, aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, cathepsin B, IGFBP-2, brain signaling protein 6C, IGF-2, and PDGF R.α, Sorting protein, Serine protease inhibitor B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, Coagulation regulator protein, Endoglycosides, IGFBP-3, RGM-C, PF4, MIF, TGM4, Periostrin, Flintase, TIMP-1, PAPP-A, Glycerin proteoglycan, PCK1, Arylsulfatase A, CD99, CA2, PRDX4, Transferrin, DcR3, GP73, LAIR2, ULBP-4, Optical proteoglycan, TIMP-2, TFPI, SOX2, SLITRK5, FAP, Spinal cord protein, ENPP-2, CD97, CTACK, Integrin α1, EXTL3, IL-18, BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin I, IGF-2R, ADAMTS L1-1, Erythropoietin, Convolutional D1, DNMT3A, BCL-2, CL-P1, Hepatocyte-B3, FABP6, CHI3L1, FCRLS, TFF3, Nephlebotomycin, DPPII, cIAP-1, PDGF Rb, Pentanoic Acid 3, Angiotensinogen, Follicle-Stapressin, CF VII, Persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Lactone, Granulosin Progenitor, PCSK2, GKN1, IL-18, Enkephalin, Stabilin-2, IL-17 RD, albumin, follicle-stimulating protein 1, MMP-10, FKBP51, LRRC4, Pref-1, galactoglobin-1, troponin C, UNC5H3, FLRT2, CD314, brain signaling protein 6B, cytokinin-4, CD27 ligand, IL-20 Rβ, brain signaling protein 6A, TSK, cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, norin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Glycopropyl peptide, Activator Protein A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Phosphatidylinositol Proteoglycan 2, MSP R, DSCAM, Membrane-type Serine Protease, KIR2DL3, CD30, Salivary Agglutinin-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAKR, Nestin-1, CD2, Kallikrein 1, TSLP R, LAMP1, TROY, VCAM-1, Salivary agglutinin-11, S100A1, PAR1, Thyroid peroxidase, Aminopeptidase P2, IL-1 RI, ADAMS, OSM R β, Platelet-Reactive Protein-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granulase B, Lipocontin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Lipoprotein, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 α, OX40 ligand, Pax3, UCH-L3, cMASP3, Langerin, desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, conglomerate A4, brain signaling protein 4D, ROBO2, PDX-1, APRIL, neuronal rank protein, transmembrane protein-2 with ring structure, EMMPRIN, activating protein RIB, neuroligand 2, epithelial regulatory protein, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, extracellular matrix protein-2, sFRP-3, p53, EphB3, NCK1, brain signaling protein 7A, NKp80, prolactin, cysteine ​​agonist protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Brain signaling protein 6D, FGF-3, GATA-4, VAP-A, CHST2, Pregnancy-associated plasma protein-2, Cohesin-3, Serpentin-1, AKR1C4, Oligosin-2, Osteoadhesin, NKp44, Thyroglobulin, IL-21R, Chemokines, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin α5. VAMP-2, FLRG, Ret metaphase factor, CD73, TRAP, proGRP, granzyme H, PRX2, p2'7, salivary lectin-6, dendritic cell-associated C-type lectin-1, CD51, Notch-1, calreticulin, DR3, DCTN1, CDC25B, bone activator, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, cathepsin V, testosterone-2, phosphatidylinositol proteoglycan 5, CD6, salivary lectin-2, asparagine endopeptidase, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 β, TAFA1, podocyte protein, RalA, CRELD2, GRAP2, SP-D, BID, GFRα-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, cathepsin L, 6-Ckine, MIS RII, kallikrein 5, TGM3, FCAR, contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, disaccharide proteoglycan, neurodevelopmental factor, ILT4, uPAR, Axl, WIF-1, IL-7 R α, GPR56, CEACAM-3, MCEMP1, FABP2, plexin B3, MEPE, activator RIIA, ANG-2, Cochlin, presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, stalkin-1, GUSB, nestin-2, IL-17F, SR-AI, TAFA2, N-cadherin, IL-17B, IL-17RC, MIP-3b, cysteine ​​protease C, cysteine ​​protease D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, intestinal peptidase, cathepsin E, TSLP, TCN2, GDF-15, epidermal morphogenetics, GRKS, PD-1, serine protease inhibitor A4, ADAM23, NOV, galactoglobulin-2, neuronal surface protein 3β, TLR3, Sirtuin 2, Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, lymphocyte chemokine, cysteine ​​protease protein SN, JAM-A, calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4Ra, kallikrein 14, extracellular matrix protein-3, Oligo2, kallikrein 12, CA13, IL-9, stalk protein-3, MPIF-1, cysteine ​​protease protein S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, transmembrane serine protease, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, brain signaling protein 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-1ra, Flatfoot protein, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenosynovin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Salivary lectin-9, ICAM-3, Cysteine ​​protease inhibitor SA, Galactohemagglutinin-4, Pepsinogen II, Desmosome core protein-3, Stalk protein-4, SCF, Serine protease inhibitor A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galactohemagglutinin-9, vWF-A2, TACE, Activator protein RIM, Cathepsin S, LDL R, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, phosphatidylinositol proteoglycan 1, LAP (TGFb1), Gash, contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, neural epithelial stem cell protein, TPO, ErbB3, Kirrel3, FLRT1, galactolectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 R α1. AMIGO2, HCC-1, TFPI-2, ULBP-2, desmosome core protein 2, agglutinin, synaptic fusion protein 4, VAMP-1, stalkin-2, FGF-21, Flt-3, GFAP, TIM-1, inhibin A, cadherin-4, PIGF-2, neurogranulin, HE4, IL-23R, galactolectin-7, GALNT3, GITR L, CD14, R-vertebral protein 2, CK19, cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, salivary lectin-7, CD155, VEGF-C, TNF-α, PGRP-S, SDF-1α, PDGF-AB, GPVI, CD40, SCF-R, platelet-reactive protein-5, IL-1 RII, neurofeltin-2, cadherin-13, E-selectin, GITR, WISP-1, renin, AgRP, MDL-1, ROBO3, RANTES, endothelial cell-specific molecules, granzyme, hCGb, mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, trypsin 3, ARSB, LIF Rα, BAFF R, CD157, granzyme A, 2B4, ESAM, IL-1R4, CXCL14, IL-31, SIRP α, urinary modulatory proteins, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3, IL-32 α, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, fibrinogen-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Synaptic Fusion Protein 6, GRO, Bcl-w, Lipocrine-2, PDGF-AA, IL-2 Ra, Angiopoietin, LYVE-1, CD4, RAGE, CDNF, Short Proteoglycan, NAP-2, PU.1, EDAR, ADAMTS13, Kynurenase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, procalcitonin, SOX15, kallikrein 11, BCMA, dendritic cell-associated C-lectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, resistin, HVEM, ENPP-7, cohesin-4, IL-2Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, brain signaling protein 4G, IL-12p40, total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, endothelial glycoprotein, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, galactolectin-8, CA4, cysteine ​​protease protein EM, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7, and IL-13. Table 1 below shows non-limiting examples of the selection of growth factors or growth factor proteins.

[0033] Table 1 In some embodiments, the therapeutic composition (e.g., an MSC secretome composition) comprises one or more nucleic acid biomolecules. At least one of the one or more nucleic acid biomolecules may be miRNA. For example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 or more nucleic acids may be present in the MSC secretome. Unrestricted example miRNAs that may be present in the MSC secretome include: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa -mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p , hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1 , hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hs a-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5 p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a,hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p, hsa-miR-376a-3p, hsa-miR-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p and hsa-miR-99b-5p.

[0034] Exemplary microRNAs that may be present in therapeutic compositions (e.g., MSC secretomes) include hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7g-5p, hsa-let-7i-5p, hsa-miR-214-3p, and hsa-miR-27a-3p, and combinations of two or more thereof, each of which has a binding site for TMPRSS2 in the mRNA.

[0035] In some embodiments, the therapeutic composition (e.g., MSC secretome) contains CD63 + CD9 - CD81 - Extracellular vesicles of the phenotype. In some embodiments, at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, or 95% of the extracellular vesicles in the therapeutic composition are CD63. + CD9 - CD81 - In some embodiments, at least 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, or 95% of the extracellular vesicles in the therapeutic composition are CD9-. In some embodiments, at least 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, or 95% of the extracellular vesicles in the therapeutic composition are CD81-. In one exemplary embodiment, the MSC is bone marrow MSC.

[0036] In some embodiments, the MSCs cultured to produce the therapeutic composition have the ability to differentiate in vitro into three lineages: adipocytes, osteoblasts, and chondrocytes. In some embodiments, the MSCs are positive for CD73, CD105, CD166, and CD90, and negative for CD14, CD31, CD34, and CD45. In one exemplary embodiment, the MSCs are bone marrow MSCs.

[0037] The therapeutic compositions described herein may comprise one or more exosomes and one or more growth factors. The growth factors and / or exosomes may be heterologous or homologous. The growth factors and / or exosomes may be derived from any cells in the human body, such as ectoderm cells, endoderm cells, or mesodermal cells. For example, the therapeutic composition may comprise growth factors derived from mesenchymal stem cells (MSCs), MSC-derived exosomes, or both MSC-derived growth factors and exosomes. In some embodiments, the therapeutic composition comprises one or more components derived from the MSC secretome and additives of components not derived from the MSC secretome. In one aspect, this article discloses a therapeutic composition comprising MSC secretome and one or more additional additives, wherein the one or more additional additives comprise prostaglandin E2 (PGE2), transforming growth factor 131 (TGF-131), hepatocyte growth factor (HGF), stromal cell-derived factor-1 (SDF-1), nitric oxide, indoleamine 2,3-dioxygenase, interleukin-4 (IL-4), IL-6, interleukin-10 (IL-10), IL-1 receptor antagonist and soluble TNF-α receptor, insulin-like growth factor, fibroblasts Growth factors (FGF) 1-23 (especially FGF1 and FGF2), bone morphogenetic protein (BMP) 1-15, epidermal growth factor (EGF), transforming growth factor-a (TGF-a), macrophage stimulating protein (MSP), platelet-derived growth factor (PLGF), vascular endothelial growth factor (VEGF), macrophage colony-stimulating factor (M-CSF), insulin, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), estrogen or thyroid hormone, or a combination of two or more thereof.

[0038] Embodiments of the therapeutic composition described herein may include proteins and microRNAs, some of which may be embedded in or surrounded by a lipid membrane to generate vesicles ranging in size from about >20 nm to about 200 nm. The number of vesicles within the composition may be about 1 million to about 100 billion vesicles / mL when in suspension, or about 10 million to about 1 trillion when formulated as a lyophilized powder. The vesicles may be extracellular vesicles. Extracellular vesicles may be components of the MSC secretome. The vesicles may be exosomes. Exosomes may be components of the MSC secretome. The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to a subject. The intravenous solution may be about 100 mL. The number of EVs in the composition can be from about 10 million to about 1 trillion. The number of EVs in the composition can be from about 10 billion to about 1 trillion.

[0039] mesenchymal stem cells The example therapeutic compositions described herein comprise MSC secretome compositions and / or components derived from mesenchymal stem cells (MSCs) (e.g., biomolecules such as nucleic acids like miRNAs, and peptides such as growth factors). In one aspect, this document discloses MSC secretome compositions (including, but not limited to, MSC growth factors, MSC exosomes, MSC extracts, and / or compositions comprising extracellular vesicles) for the treatment, inhibition, mitigation, reduction, improvement, and / or prevention of conditions such as, for example, graft-versus-host disease.

[0040] MSCs are pluripotent cells capable of differentiating into multiple cell types, including myocytes, chondrocytes, adipocytes, and osteoblasts. These cells are typically found in the placenta, umbilical cord blood, adipose tissue, bone marrow, or amniotic fluid, including perivascular tissue. As used herein, “MSC” refers to non-terminally differentiated cells, including but not limited to pluripotent stem cells, pluripotent stromal cells, stromal vascular cells, pericytes, vascular pericytes, stromal cells, pluripotent cells, fibroblast-like cells derived from adipose tissue, stromal vascular fractions derived from adipose tissue, MSCs derived from bone marrow, fibroblast-like cells derived from bone marrow, stromal vascular fractions derived from bone marrow, MSCs derived from bone marrow, fibroblast-like cells derived from tissue, adult stem cells, adult stromal cells, keratinocytes, and / or melanocytes.

[0041] In some embodiments, the MSC secretome composition comprises MSC growth factors, MSC exosomes, extracellular vesicles, extracellular vesicle isolates (EVIPs), cell-free extracts of MSCs, or MSC lysates, or combinations of two or more thereof, derived from MSCs. MSCs can be human MSCs, fibroblast-like cells, or non-human animal MSCs, including but not limited to MSCs from horses, cattle, pigs, sheep, non-human primates, dogs, cats, rabbits, rats, or mice. In embodiments, MSCs can be derived from the patient to whom the composition will be applied (autologous) or from another individual (allogeneic). MSCs can be cultured and expanded to collect conditioned medium and / or increase the cell count of the lysates, or used fresh before incorporation into the therapeutic composition of this disclosure.

[0042] MSC secretome compositions (including, but not limited to, MSC growth factors, MSC exosomes, MSC extracts and / or compositions containing extracellular vesicles) may contain about 0.00001 to about 20 wt.%, such as about 0.01 to about 10 wt.% of mesenchymal stem cell (MSC) extracts, MSC exosomes or MSC growth factor formulations.

[0043] The MSC secretome composition or its components can be obtained by culturing MSCs under normal hyperoxia conditions or under artificial wound healing conditions.

[0044] MSCs can be selectively stimulated to produce the MSC secretome compositions or components thereof disclosed herein. Stimulated MSCs can produce MSC growth factors, secretomes, cytokines, chemokines, mesenchymal stem cell proteins, peptides, glycosaminoglycans, extracellular matrix (ECM), proteoglycans, or extracellular vesicles (e.g., exosomes), or combinations of two or more thereof. MSC growth factors include, but are not limited to, prostaglandin E2 (PGE2), transforming growth factor 131 (TGF-131), hepatocyte growth factor (HGF), stromal cell-derived factor-1 (SDF-1), nitric oxide, indoleamine 2,3-dioxygenase, interleukin-4 (IL-4), IL-6, interleukin-10 (IL-10), IL-1 receptor antagonists and soluble TNF-α receptors, insulin-like growth factor, fibroblast growth factor (FGF) 1-23 (especially FGF1 and FGF2), bone morphogenetic protein (BMP) 1-15, epidermal growth factor (EGF), transforming growth factor-a (TGF-a), macrophage-stimulating protein (MSP), platelet-derived growth factor (PLGF), vascular endothelial growth factor (VEGF), macrophage colony-stimulating factor (M-CSF), insulin, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and hormones, including estrogen and thyroid hormones.

[0045] MSC culture can occur under wound healing and / or hypoxic conditions. Wound healing conditions can include approximately 1% to approximately 5% oxygen, reduced or absent serum, reduced glucose, or various combinations of these elements. Combined reduced nutrient and metabolite environments can trigger cultured cells to produce wound-healing and anti-inflammatory ECM proteins and growth factors to guide tissue healing. In one aspect, the MSC secretome composition comprises MSC growth factors, MSC exosomes, and / or MSC cell extracts or MSC lysates obtained from MSCs cultured under standard hyperoxia conditions (e.g., 21% oxygen) or under artificial wound healing conditions (e.g., such as 0.1% to approximately 5% oxygen).

[0046] Artificial wound healing conditions for culturing MSCs may include one or more of the following growth conditions: reduced glucose availability, reduced oxygen tension, decreased pH, and increased temperature.

[0047] In some embodiments, glucose availability may be reduced relative to a normal control (e.g., 4.5 g / L). The glucose in modified culture media that reduce glucose without harming cells may be reduced by 0% to 50%, more preferably by about 5% to 40%. For example, MSC artificial wound healing culture conditions may include glucose reductions of about 5% to about 15%, about 10% to about 20%, about 15% to about 25%, about 20% to about 30%, or about 25% to about 35%. In some embodiments, glucose is present at a concentration of about 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, or 4.0 g / L, or at a concentration in the range of any two of these values. In some embodiments, glucose is present at a concentration of less than or no more than 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, or 4.5 g / L.

[0048] In some implementations, oxygen tension can be reduced to the oxygen levels of hypoxic conditions. Normal atmospheric oxygen is approximately 21%, and any reduction is considered hypoxic. Therefore, in one aspect, MSCs can be cultured at oxygen levels of 0.0% to 20.9%, from about 0.1% to about 0.5%, from about 0.1% to about 2.0%, from about 0.1% to about 5.0%, from about 0.5% to 5.0%, from about 1.0% to about 10%, from about 5.0% to about 10.0%, and from about 10.0% to about 15.0%. Hypoxic conditions can be one aspect of artificial wound healing conditions.When MSCs are cultured to produce a therapeutic secretory composition containing extracellular vesicles and / or growth factors secreted by MSCs, the oxygen tension can be between about 0.5% and 20.5% oxygen, for example, such as 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4 5, 4.6, 4.7, 4.8, 4.9, 5, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7, 10.8, 10.9, 11, 11.1, 11.2, 11.3, 11.4, 11.5, 11.6, 11.7, 11.8, 11.9, 12, 12.1, 12.2, 12.3, 12.4, 12.5, 12.6, 12.7, 12.8, 12.9, 13, 13.1, 13.2, 13.3, 13.4, 13.5, 13.6, 13.7, 13.8, 13.9, 14, 14.1, 14.2, 14.3, 14.4, 14.5, 14.6, 14.7, 14.8, 14.9, 15, 15.1, 15.2, 15.3, 15.4, 15 0.5, 15.6, 15.7, 15.8, 15.9, 16, 16.1, 16.2, 16.3, 16.4, 16.5, 16.6, 16.7, 16.8, 16.9, 17, 17.1, 17.2, 17.3, 17.4, 17.5, 17.6, 17.7, 17.8, 17.9, 18, 18.1, 18.2, 18.3, 18.4, 18.5, 18.6, 18.7, 18.8, 18.9, 19, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, or 20.0% oxygen, or a range between any two of these values.

[0049] The pH value can also be lowered during MSC culture. The pH can be from about 6.0 to about 7.4, for example, from about 6.0 to about 6.4, from about 6.2 to about 6.4, from about 6.2 to about 6.6, from about 6.4 to about 6.6, from about 6.4 to about 6.8, or from about 6.6 to about 7.0, such as 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, or 7.4.

[0050] The temperature of the culture environment can be increased relative to the physiological homeostatic temperature (e.g., 37°C). In one aspect, the culture conditions for MSCs can include from about 35°C to about 39°C, from about 35°C to about 36°C, from about 36°C to about 37°C, from about 37°C to about 38°C, from about 38°C to about 39°C, and from about 39°C to about 40°C. In another aspect, the culture temperature can be 35.0, 35.1, 35.2, 35.3, 36.4, 35.5, 35.6, 35.7, 35.8, 35.9, 36.0, 36.1, 36.2, 36.3, 36.4, 36.5, 36.6, 36.7, 36.8, 36.9, 37.0, 37.1, 37.2, 37.3, 3... 7.4, 37.5, 37.6, 37.7, 37.8, 37.9, 38.0, 38.1, 38.2, 38.3, 38.4, 38.5, 38.6, 38.7, 38.8, 38.9, 39.0, 39.1, 39.2, 39.3, 39.4, 39.5, 39.6, 39.7, 39.8, 39.9 or 40.0℃.

[0051] In some embodiments, the culture medium is serum-free. In some embodiments, the serum-free culture medium contains platelet lysate. In some embodiments, the platelet lysate is human platelet lysate (HPL). In some embodiments, the serum-free culture medium contains at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% HPL by volume, or any range between two of these values. In some embodiments, the culture medium contains from 8% to 12%, 5% to 15%, or 9% to 11% HPL by volume.

[0052] In one aspect, therapeutic compositions, such as MSC secretome compositions (including but not limited to MSC growth factors, MSC exosomes, MSC extracts, and / or compositions containing extracellular vesicles), contain additives. Additives can act as a protective coating. Additives can reduce the degradation of components within the composition, such as growth factors. Additives can be cryoprotectants. Cryoprotectants can be oligosaccharides. Additives can be polymers. Polymers include relatively high molecular weight organic compounds, natural or synthetic, whose structure can be represented by repeating small unit monomers. Non-limiting examples of polymers include polyethylene, rubber, and cellulose. Synthetic polymers are typically formed by the addition or condensation polymerization of monomers. The term "copolymer" can refer to a polymer formed from two or more different repeating units (monomer residues). For example, but not limited to, copolymers can be alternating copolymers, random copolymers, block copolymers, or graft copolymers. Polymers can be natural polymers, synthetic polymers, homopolymers, heteropolymers, or copolymers. In one aspect, polymers can comprise copolymers, block copolymers, diblock copolymers, and / or triblock copolymers. In one aspect, additives can comprise biocompatible polymers. In one aspect, biocompatible polymers are crosslinked. Biocompatible polymers include, but are not limited to, polysaccharides; hydrophilic peptides; polyamino acids, such as poly-L-glutamic acid (PGS), γ-polyglutamic acid, poly-L-aspartic acid, poly-L-serine, or poly-L-lysine; polyalkylene glycols and polyalkylene oxides, such as polyethylene glycol (PEG), polypropylene glycol (PPG), and polyethylene oxide (PEO); poly(oxyethylated polyols); poly(enols); polyvinylpyrrolidone; poly(hydroxyalkyl methacrylamide); poly(hydroxyalkyl methacrylate); polysaccharides; poly(hydroxy acids); poly(vinyl alcohol), polyhydroxy acids such as poly(lactic acid), poly(glycolic acid), and polylactic-glycolic acid copolymers; and polyhydroxyalkanoates. Esters such as poly(3-hydroxybutyrate) or poly(4-hydroxybutyrate); polycaprolactone; poly(orthoester); polyanhydride; poly(phosphazene); polylactide-caprolactone copolymer; polycarbonates such as tyrosine polycarbonate; polyamides (including synthetic and natural polyamides), peptides and poly(amino acids); polyesteramide; polyester; poly(p-dioxanone); poly(alkylene oxide); hydrophobic polyether; polyurethane; polyether ester; polyacetal; polycyanoacrylate; polyacrylate; polymethyl methacrylate; polysiloxane; poly(ethylene oxide) / poly(propylene oxide) copolymer; polyketide; polyphosphate; polyhydroxyvalerate; polyalkylene oxalate; polyalkylene succinate; poly(maleic acid) and its copolymers.Biocompatible polymers may also include polyamides, polycarbonates, polyalkylenes, polyalkylene glycols, polyalkylene oxides, polyalkylene terephthalates, polyvinyl alcohol (PVA), methacrylate PVA (m-PVA), polyethylene ethers, polyethylene esters, polyhalogenated polyethylene, polyvinylpyrrolidone, polyglycolic acid, polysiloxanes, polyurethanes and their copolymers, alkyl cellulose, hydroxyalkyl cellulose, cellulose ethers, cellulose esters, nitrocellulose, polymers of acrylates and methacrylates, methylcellulose, ethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, hydroxybutyl methylcellulose, cellulose acetate, cellulose propionate, cellulose acetate butyrate, cellulose acetate phthalate, carboxyethylcellulose, cellulose triacetate, sodium cellulose sulfate, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), and poly(isobutyl methacrylate). Poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate), polyethylene, polypropylene, poly(ethylene glycol), poly(ethylene oxide), poly(ethylene glycol terephthalate), poly(vinyl alcohol), poly(vinyl acetate, polyvinyl chloride, polystyrene, and polyvinylpyrrolidone), their derivatives, their linear and branched copolymers and block copolymers, and blends thereof. Exemplary biodegradable polymers include polyesters, poly(orthoesters), poly(ethyleneamine), poly(caprolactone), poly(hydroxybutyrate), poly(hydroxyvalerate), polyanhydride, poly(acrylic acid), polyglycolic acid, poly(glycolic acid), poly(urethane), polycarbonate, polyphosphate, polyphosphazene, their derivatives, their linear and branched copolymers and block copolymers, and mixtures thereof.

[0053] In some embodiments, the protective coating comprises a carbohydrate structure of monosaccharides and carbohydrate polymers (such as disaccharides or polysaccharides, including but not limited to non-reducing polysaccharides or disaccharides and any combination thereof). Examples of carbohydrates that can be used in the protective coating include glucose, aldoses (D-allose, D-agrose, D-mannose, etc.), glucopyranose, pentahydroxyhexanal, aD-glucopyranosyl-D-glucose, aD-glucopyranosyl-dihydrate, polymers of PD-glucopyranosyl units, PD-fructofuranosyl-aD-glucopyranoside (anhydrous / dihydrate), f3-D-galactopyranosyl-D-glucose, aD-glucopyranosyl-dihydrate, etc. Polymers of glycosyl-aD-glucopyranoside (anhydrous / dihydrate), galactose, pentoses (ribose, xylose, lysose), dextrose, decahydrate, fructose, sucrose, lactose, maltose, trehalose, agarose, D-galactosyl-O-(1-4)-dehydrated-L-galactosyl, cellulose, PD-glucopyranoside units, and starch, as well as polyols, polyols, sugar alcohols, erythritol, glycols, glycerol, xylitol, and sorbitol.

[0054] In some embodiments, the protective coating contains a biocompatible and / or biodegradable polyester or polyanhydride, such as poly(lactic-co-glycolic acid), poly(glycolic acid), and poly(lactic-co-glycolic acid) copolymers. The particles may comprise one or more of the following polyesters: homopolymers comprising units of glycolic acid (hereinafter referred to as “PGA”), and lactic acid units (such as poly-L-lactic acid, poly-D-lactic acid, poly-D,L-lactic acid, poly-L-lactide, poly-D-lactide, and poly-D,L-lactide 5, collectively referred to as “PLA”), and caprolactone units (such as poly(e-caprolactone) (collectively referred to as “PCL”); copolymers comprising lactic acid and glycolic acid units, such as various forms of polylactic-co-glycolic acid copolymers and polylactide-glycolic acid copolymers, characterized by a lactic acid:glycolic acid ratio, collectively referred to as “PLGA”); and polyacrylates and their derivatives. Exemplary polymers also include copolymers of polyethylene glycol (PEG) and the aforementioned polyesters, such as various forms of PLGA-PEG or PLA-PEG copolymers, collectively referred to herein as "PEGylated polymers". In some embodiments, the PEG regions can be covalently associated with the polymer via a cleavable joint to create a "PEGylated polymer". In one aspect, the polymer comprises at least 60%, 65%, 70%, 75%, 80%, 85%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% acetal side groups.

[0055] The triblock copolymers disclosed herein include, for example, polyethylene glycol (PEG), polyvinyl acetate, polyvinyl alcohol, polyvinylpyrrolidone (PVP), polyethylene oxide (PEO), polyvinylpyrrolidone-vinyl acetate copolymer, polymethacrylate, polyoxyethylene alkyl ether, polyoxyethylene castor oil, polycaprolactam, polylactic acid, polyglycolic acid, poly(lactic-glycolic acid), polylactic-glycolic acid copolymer (PLGA), and cellulose derivatives such as hydroxymethyl cellulose and hydroxypropyl cellulose. Examples of diblock copolymers that can be used in the protective coatings disclosed herein include, for example, polyethylene glycol (PEG), polyvinyl acetate, polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyethylene oxide (PEO), polyvinylpyrrolidone-vinyl acetate copolymer, polymethacrylate, polyoxyethylene alkyl ether, polyoxyethylene castor oil, polycaprolactam, polylactic acid, polyglycolic acid, poly(lactic-glycolic acid), polylactic-glycolic acid copolymer (PLGA).

[0056] In one aspect, the protective coating comprises (i.e., encapsulated), and the encapsulated composition may further comprise lecithin or hydrolyzed lecithin as a carrier or as an encapsulating material. As used herein, lecithin and / or hydrolyzed lecithin coatings comprise coatings containing phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine, phosphatidylserine, and phosphatidic acid. The source of lecithin may be plant or animal.

[0057] In one aspect, any of the polymers, monosaccharides, disaccharides, or polysaccharides used to form the protective coating formed by placing the MSC additive in the encapsulation solution can be at an appropriate concentration for forming the protective coating. For example, the polymers, monosaccharides, disaccharides, or polysaccharides can be at any concentration between 0.01 mM and 10.0 M, such as from about 0.01 M to about 0.1 M, from about 0.1 mM to about 1.0 M, or from about 1.0 M to about 10.0 M.

[0058] In one aspect, the MSC secretome compositions disclosed herein (including, but not limited to, MSC growth factors, MSC exosomes, MSC extracts, and / or compositions comprising extracellular vesicles) may include any known ingredients commonly found in the pharmaceutical field, such as agents for combating free radicals; bactericides; isolating agents; preservatives; alkalizing or acidifying agents; fragrances; surfactants; fillers; natural products or extracts of natural products, such as aloe or green tea extracts; vitamins; or coloring materials. Other ingredients that may be combined with the powder may include antioxidants, which may be selected from a variety of antioxidants. Suitable antioxidants include vitamins such as vitamin C (L-ascorbic acid, magnesium ascorbate-2-phosphate, ascorbate palmitate, tetrahexyldecyl ascorbate), vitamin E (tocotrienols), vitamin A (retinol, retinal, retinoic acid, provitamin A carotenoids, such as β-carotene), N-acetylglucosamine, or other derivatives of glucosamine. Other components may include at least one essential fatty acid, such as S2-3, S2-6, and S2-9 polyunsaturated fatty acids, such as linoleic acid (LA), gamma-linolenic acid (GLA), alpha-linolenic acid (ALA), di-homo-gamma-linolenic acid (DGLA), and arachidonic acid (ARA). Fatty acids can be derived from a variety of sources, including evening primrose oil, blackcurrant oil, borage oil, or GLA-modified safflower seed oil. Other components may include a platelet-rich fibrin matrix, at least one component that supports ECM production and hyaluronic acid production, such as N-acetylglucosamine or other derivatives of glucosamine, ultra-low molecular weight (ULMW) hyaluronic acid, chondroitin sulfate, or keratin sulfate.

[0059] MSC secretome compositions can be produced by a method comprising culturing MSCs collected from donors under stimulating conditions; collecting the secretome and combining the secretome with an additive; and freezing the mixture; wherein the mixture is formulated into a therapeutic composition. The frozen mixture may be a dry powder. The therapeutic composition may comprise a dry powder reconstituted into a delivery medium (e.g., saline) suitable for human administration. The therapeutic composition may comprise exosomes, peptides, proteins, cytokines, growth factors, extracellular matrix (ECM), proteoglycans, glycosaminoglycans, chemokines, or combinations of two or more thereof. MSCs may be bone marrow MSCs. MSCs may be human MSCs, animal MSCs, pluripotent stromal cells, fibroblasts, or fibroblast cells. MSC secretome compositions can be produced by a method comprising culturing MSCs collected from donors under stimulating conditions, lysing the MSCs to produce an extracted lysate, concentrating the extracted lysate, combining the extracted lysate with an additive, and freezing the mixture to produce a dry powder. In an exemplary embodiment, the additive is a sugar. In some embodiments, freezing includes lyophilization. The powder may contain a concentrated collection of analgesic MSC secretory components (e.g., exosomes, extracellular matrix) that are specific for anti-inflammatory purposes.

[0060] The method may also include filtration-sterilization, concentration, freezing, or freeze-drying of MSC culture media, for example, after culture under wound healing conditions. Furthermore, MSC culture media may be combined with a cryoprotectant and then frozen. The cryoprotectant may be an additive, such as oligosaccharides.

[0061] Lysis can be achieved by adding a hypotonic solution or by repeated freeze-thaw cycles that disrupt the cell membrane. Cells can lyse while attached to a culture surface or in suspension. Cells can be lysed by enzymatic release and / or by mechanical homogenization.

[0062] Stimulation conditions include stimulating MSCs to selectively secrete desired anti-inflammatory proteins, peptides, glycosaminoglycans, proteoglycans, exosomes, and / or secretomes. Stimulation can be achieved by adjusting cell growth conditions such as cell confluence, culture medium supplements, nutrient supplements, oxygen levels, culture duration under these conditions, cell passage number, or combinations thereof. Stimulation can include growth under wound healing conditions. Wound healing conditions can include approximately 1% to approximately 5% oxygen, reduced or absent serum, reduced glucose, or various combinations of these elements.

[0063] Delivery of drug carriers / drug products Therapeutic compositions, such as MSC secretome compositions, can be administered in vivo in pharmaceutically acceptable carriers. "Pharmaceutically acceptable" means that the material is not biologically or otherwise undesirable; that is, the material can be administered to the subject together with the nucleic acid or carrier without causing any undesirable biological effects or interacting harmfully with any of the other components of the pharmaceutical composition containing it. The carrier can be selected to minimize any degradation of the active ingredient and any adverse side effects on the subject. The composition can be administered parenterally (e.g., intravenously).

[0064] If the composition is used for parenteral administration, it is typically characterized by injection. Injectable formulations can be prepared in conventional forms, such as liquid solutions or suspensions, solid forms suitable for dissolving or suspending in a liquid prior to injection, or emulsions. Parenteral administration may involve the use of sustained-release or continuous-release systems to maintain a constant dose.

[0065] Preparations intended for parenteral administration may include sodium chloride solution.

[0066] Therapeutic uses This article provides a method for treating graft-versus-host disease in subjects who may have received organ transplants or allogeneic grafts. Subjects may have received modified multi-organ allogeneic grafts, single intestinal allogeneic grafts, or single liver allogeneic grafts. Subjects may exhibit one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection. For example, subjects may exhibit one or more signs or symptoms selected from the following: crypt cell apoptosis, crypt loss, ulceration, allogeneic graft cell rejection, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulcers, and combinations thereof.

[0067] In some implementations, the subject may be receiving corticosteroids. For example, the subject may be receiving high-dose corticosteroids. In some implementations, the subject may experience improvement in one or more of the following signs or symptoms after treatment: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, or combinations thereof.

[0068] Methods of treating graft-versus-host disease in a subject may include administering to the subject a composition comprising a protein, wherein the protein comprises one or more proteins selected from the following: ferritin, IGFBP-4 (insulin-like growth factor binding protein-4), IL-1 R6 (interleukin-1 receptor 6), LAMP2 (lysosome-associated membrane glycoprotein 2), bIG-H3 (transforming growth factor-β-inducible protein ig-h3), GPR115 (adhesion G protein-coupled receptor F4), CD63 antigen, CD109 antigen, serine protease inhibitor F1 (pigment epithelial-derived factor), IGFBP-6 (insulin-like growth factor binding protein-6), HS3ST4 (heparanol sulfate glucosamine 3-O-sulfotransferase 4), OPN (osteopontin), PAI-1 (plasminogen activator inhibitor-1 or sERPINE). 1) Cathepsin B, IGFBP-2 (insulin-like growth factor binding protein-2), brain signaling protein 6C, IGF-2 (insulin-like growth factor-2), sorting protein, serine protease inhibitor B6, Dkk-3 (Dickkopf-related protein 3), CNTF (ciliary neurotrophic factor), TSP-1 (thromboretin-1), GM-CSF Ra (granulocyte-macrophage colony-stimulating factor receptor subunit α), coagulation regulatory protein, endosaccharide, (podocyte protein-like protein 2) IGFBP-3 (insulin-like binding protein-3), RGM-C (hepcidin regulatory protein), PF4 (platelet factor 4), MIF (macrophage migration inhibitory factor), TGM4 (protein-glutamine-γ-glutamyltransferase 4), periosteal protein, furin protease, TIMP-1 (MMP tissue inhibitor 1), gypsum glycan, PCK1 (cytosol-type phosphoenolpyruvate carboxykinase), CD9 antigen, CD99 antigen, CA2 (carbonic anhydrase 2), PRDX4 (peroxide reductase-4), transferrin, DcR3 (tumor necrosis factor receptor superfamily member 6B), GP73 (Golgi membrane protein 1), CD81 antigen, luminescent glycan, TIMP-2 (MMP tissue inhibitor 2), and the proteins listed in Table 1.Methods for treating graft-versus-host disease in a subject may include administering a composition of extracellular vesicles (EVs) to the subject, the EVs comprising one or more nucleic acids selected from the following: hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-21-5p, hsa-miR-221-3p, hsa-miR-222-3p, hsa-miR-222-3p, hsa-miR-223p, hsa-mi R-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, and hsa-miR-940, and combinations of two or more thereof. In some methods, at least 80% of the EVs are CD63+CD9-CD81-. The composition can have an EV concentration of at least about 60 billion EVs. The composition can have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution may be about 100 mL. The number of EVs in the composition may be from about 10 million to about 1 trillion. The number of EVs in the composition may be from about 10 billion to about 1 trillion.

[0069] Methods for treating graft-versus-host disease in a subject may include administering to the subject a composition comprising extracellular vesicles (EVs), wherein the composition comprises at least about 60 billion EVs. At least about 60 billion EVs can be from about 60 billion to about 80 billion EVs. At least about 60 billion EVs can be a composition of at least about 60 billion EVs / mL. At least about 60 billion EVs can be a composition of about 60 billion to 80 billion EVs / mL. In some methods, at least 80% of the EVs are CD63. + CD9 - CD81 -The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution may be about 100 mL. The number of EVs in the composition may be about 10 million to about 1 trillion. The number of EVs in the composition may be about 1 trillion. The number of EVs in the composition may be about 10 billion to about 1 trillion.

[0070] Methods for treating graft-versus-host disease in a subject may include administering to the subject a composition comprising extracellular vesicles (EVs), wherein at least 80% of the EVs are CD63. + CD9 - CD81 - The composition may have an EV concentration of at least about 60 billion EVs. The composition may have an EV concentration of at least about 60 billion EVs / mL. The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution may be about 100 mL. The number of EVs in the composition may be about 10 million to about 1 trillion. The number of EVs in the composition may be about 1 trillion. The number of EVs in the composition may be about 10 billion to about 1 trillion.

[0071] Methods for treating graft-versus-host disease in a subject may include administering to the subject a composition comprising a protein and extracellular vesicles (EVs), wherein the protein comprises one or more proteins selected from the following: ferritin, IGFBP-4 (insulin-like growth factor binding protein-4), IL-1 R6 (interleukin-1 receptor 6), LAMP2 (lysosome-associated membrane glycoprotein 2), bIG-H3 (transforming growth factor-β inducible protein ig-h3), GPR115 (adhesion G protein-coupled receptor F4), CD109 antigen, serine protease inhibitor F1 (pigment epithelial-derived factor), IGFBP-6 (insulin-like growth factor binding protein-6), HS3ST4 (heparanol sulfate glucosamine 3-O-sulfotransferase 4), OPN (osteopontin), PAI-1 (plasminogen activator inhibitor-1 or sERPINE). 1) Cathepsin B, IGFBP-2 (insulin-like growth factor binding protein-2), brain signaling protein 6C, IGF-2 (insulin-like growth factor-2), sorting protein, serine protease inhibitor B6, Dkk-3 (Dickkopf-related protein 3), CNTF (ciliary neurotrophic factor), TSP-1 (thromboretin-1), GM-CSF Ra (granulocyte-macrophage colony-stimulating factor receptor subunit α), coagulation regulatory protein, endosaccharide, (podocyte protein-like protein 2) IGFBP-3 (insulin-like binding protein-3), RGM-C (hepcidin regulatory protein), PF4 (platelet factor 4), MIF (macrophage migration inhibitory factor), TGM4 (protein-glutamine-γ-glutamyltransferase 4), periosteal protein, furin protease, TIMP-1 (MMP tissue inhibitor 1), gypsum proteoglycan, PCK1 (cytosol-type phosphoenolpyruvate carboxykinase), CD9 antigen, CD99 antigen, CA2 (carbonic anhydrase 2), PRDX4 (peroxide reductase-4), transferrin, DcR3 (tumor necrosis factor receptor superfamily member 6B), GP73 (Golgi membrane protein 1), CD81 antigen, luminescent proteoglycan, TIMP-2 (MMP tissue inhibitor 2), and the proteins listed in Table 1;Furthermore, the EV contains one or more nucleic acids selected from the following: hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-21-5p, hsa-miR-221-3p, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR- The miRNAs are hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, and hsa-miR-940, and combinations of two or more thereof. The miRNA sequences are available from https: / / www.mirbase.org / . In some methods, at least 80% of the EVs are CD63+CD9-CD81-. The composition can have an EV concentration of at least approximately 60 billion EVs / mL. The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution may be about 100 mL. The number of EVs in the composition may be about 10 million to about 1 trillion. The number of EVs in the composition may be about 1 trillion. The number of EVs in the composition may be about 10 billion to about 1 trillion.

[0072] The protein administered to the subject can be prepared by a method comprising: (a) culturing bone marrow-derived mesenchymal stem cells (MSCs) under conditions of: (i) oxygen tension below 5%; and (ii) a pH below 7; (b) harvesting the MSC conditioned medium; and (c) formulating the MSC conditioned medium to produce a composition comprising the protein produced from the bone marrow-derived MSCs in step (a), and the formulation comprising formulating the protein into a pharmaceutically acceptable formulation for injection into the subject. The medium may be serum-free. The medium may have a glucose concentration of less than 4.5 g / L or be glucose-free. The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution can be approximately 100 mL. The number of EVs in the composition can be approximately 10 million to approximately 1 trillion. The number of EVs in the composition can be approximately 1 trillion. The number of EVs in the composition can be approximately 10 billion to approximately 1 trillion.

[0073] The EVs administered to the subject can be prepared by a method comprising: (a) culturing bone marrow-derived mesenchymal stem cells (MSCs) under the following conditions to produce an MSC conditioned medium: (i) oxygen tension below 5%; and (ii) a medium with a pH below 7; (b) harvesting the MSC conditioned medium; and (c) preparing the MSC conditioned medium to produce a composition comprising the EVs produced from the bone marrow-derived MSCs in step (a), and the preparation comprising formulating the EVs into a pharmaceutically acceptable formulation for injection into the subject. The medium may be serum-free. The medium may have a glucose concentration below 4.5 g / L or be glucose-free. In some methods, at least 80% of the EVs are CD63. + CD9 - CD81 -The composition may have an EV concentration of at least about 60 billion EVs. The composition may have an EV concentration of at least about 60 billion EVs / mL. The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL of the composition is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution may be about 100 mL. The number of EVs in the composition may be about 10 million to about 1 trillion. The number of EVs in the composition may be about 1 trillion. The number of EVs in the composition may be about 10 billion to about 1 trillion.

[0074] A method for treating graft-versus-host disease in a subject may include administering a protein-containing composition to the subject, the method comprising: (a) culturing bone marrow-derived mesenchymal stem cells (MSCs) under conditions of: (i) oxygen tension below 5%; and (ii) a pH below 7; (b) harvesting the MSC conditioned medium; and (c) preparing the MSC conditioned medium to produce a composition comprising the protein produced from the bone marrow-derived MSCs in step (a), and the preparation comprising formulating the protein into a pharmaceutically acceptable formulation for injection into the subject; and (d) administering the pharmaceutically acceptable formulation to the subject. The culture medium may be serum-free. The culture medium may have a glucose concentration of less than 4.5 g / L or may be glucose-free.

[0075] A method for treating graft-versus-host disease in a subject may include administering a composition comprising EVs to the subject, the method comprising: (a) culturing bone marrow-derived mesenchymal stem cells (MSCs) to produce an MSC conditioned medium under conditions of: (i) oxygen tension below 5%; and (ii) a pH below 7; (b) harvesting the MSC conditioned medium; and (c) preparing the MSC conditioned medium to produce a composition comprising EVs produced from the bone marrow-derived MSCs in step (a), and preparation comprising formulating the EVs into a pharmaceutically acceptable formulation for injection into the subject; and (d) administering the pharmaceutically acceptable formulation to the subject. The medium may be serum-free. The medium may have a glucose concentration of less than 4.5 g / L or be glucose-free. In some methods, at least 80% of the EVs are CD63. + CD9 - CD81 -The composition may have an EV concentration of at least about 60 billion EVs. The composition may have an EV concentration of at least about 60 billion EVs / mL. The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL of the composition is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution may be about 100 mL. The number of EVs in the composition may be about 10 million to about 1 trillion. The number of EVs in the composition may be about 1 trillion. The number of EVs in the composition may be about 10 billion to about 1 trillion.

[0076] Methods for treating graft-versus-host disease in a subject may include administering a composition comprising a protein and an EV to the subject, the method comprising: (a) culturing bone marrow-derived mesenchymal stem cells (MSCs) to produce an MSC conditioned medium under conditions of: (i) oxygen tension below 5%; and (ii) a pH below 7; (b) harvesting the MSC conditioned medium; and (c) preparing the MSC conditioned medium to produce a composition comprising a protein and an EV produced from the bone marrow-derived MSCs in step (a), and preparation comprising formulating the protein and EV into a pharmaceutically acceptable formulation for injection into the subject; and (d) administering the pharmaceutically acceptable formulation to the subject. The medium may be serum-free. The medium may have a glucose concentration of less than 4.5 g / L or be glucose-free. In some methods, at least 80% of the EVs are CD63. + CD9 - CD81 - The composition may have an EV concentration of at least about 60 billion EVs. The composition may have an EV concentration of at least about 60 billion EVs / mL. The composition may have an EV concentration of at least about 60 billion EVs / mL, for example, about 60 billion to about 250 billion EVs / mL, or about 60 billion to about 80 billion EVs / mL. Optionally, about 5, 10, 15, or 20 mL of the composition is administered. As a non-limiting example, about 5, 10, 15, or 20 mL of the composition is diluted in an intravenous solution (such as saline) and administered to the subject. The intravenous solution may be about 100 mL. The number of EVs in the composition may be about 10 million to about 1 trillion. The number of EVs in the composition may be about 1 trillion. The number of EVs in the composition may be about 10 billion to about 1 trillion.

[0077] In some embodiments, the therapeutic compositions described herein (such as MSC secretome compositions) comprise growth factors and / or exosomes. Growth factors and / or exosomes can induce cell proliferation and angiogenesis. The therapeutic compositions described herein may comprise mitogenic proteins such as transforming growth factor-α (TGF-α), TGFβ, hepatocyte growth factor (HGF), epidermal growth factor (EGF), basic fibroblast growth factor-2 (FGF-2), and / or insulin-like growth factor-1 (IGF-1). These increase the division of fibroblasts, epithelial cells, and endothelial cells. The therapeutic compositions described herein may comprise vascular endothelial growth factor (VEGF), IGF-1, EGF, and / or angiopoietin-1, which recruit endothelial lineage cells and initiate vascularization. The therapeutic compositions described herein may comprise anti-inflammatory components. The therapeutic compositions described herein may comprise immunomodulatory components. Non-limiting exemplary anti-inflammatory proteins include prostaglandin 2, TGF-131, HGF, SDF-1, nitrous oxide, indoleamine 2,3-dioxygenase, IL-4, IL-10, IL-1 receptor antagonists, and soluble tumor necrosis factor-α receptors. The therapeutic compositions of this invention may contain components that inhibit the proliferation and function of inflammatory immune cells, including T cells, natural killer cells, B cells, monocytes, macrophages, and dendritic cells.

[0078] A chronic inflammatory environment is characterized by a persistent imbalance between helper T cell types and macrophage types. In some implementations, therapeutic compositions, such as MSC secretory compositions, indirectly promote the conversion of TH1 cells to TH2 cells by reducing INF-γ and increasing IL-4 and IL-10. The restored TH1 / TH2 balance can improve tissue regeneration in cartilage, muscle, and other soft tissues. The reduction of INF-γ and the secretion of IL-4 can promote the conversion of macrophages from M1 type (pro-inflammatory, anti-angiogenic, and tissue growth inhibitory) to M2 type (anti-inflammatory, pro-remodeling, and tissue healing) for healing and regeneration of bone, muscle, and nerves.

[0079] Treatment methods for graft-versus-host disease (GVHD) In some embodiments, the therapeutic compositions disclosed herein (e.g., MSC secretome compositions) are used in methods of treating a subject with graft-versus-host disease (GVHD). Any therapeutic composition described herein can be used in such methods. In some embodiments, the subject is taking an immunosuppressive drug. In some embodiments, the subject has a modified multi-organ allogeneic graft. In some embodiments, the subject has a separate intestinal allogeneic graft. In some embodiments, the subject has a separate liver allogeneic graft. In some embodiments, the subject exhibits signs or symptoms of graft-versus-host disease. In some embodiments, the subject exhibits signs or symptoms of allogeneic graft rejection. In some implementations, signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection include, but are not limited to, crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, and / or ulceration.

[0080] In some embodiments, the subject is receiving a high dose of corticosteroids. In some embodiments, despite receiving a high dose of corticosteroids, the subject exhibits signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection. In some embodiments, the subject shows improvement in one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection after administration of one or more doses of the treatment product. In some embodiments, the subject shows improvement within 24 hours of administration of the treatment product. In some embodiments, the subject shows improvement in pulmonary symptoms and / or dermatological manifestations of graft-versus-host disease after administration of the treatment product. In some embodiments, the subject shows improvement in pulmonary symptoms and / or dermatological manifestations of graft-versus-host disease within 24 hours of administration of the treatment product. In some embodiments, the subject shows improvement in one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection within 7 days of administration of the initial dose of the treatment product. In some embodiments, the subject shows improvement in serological laboratory assessments related to graft function within 7 days of administration of the initial treatment dose. In some implementations, the subject exhibits complete histological resolution of graft inflammation and / or rejection within 7 days of application of the treatment product.

[0081] In some embodiments, the treatment product is administered intravenously. In some embodiments, 15 mL of the treatment product is administered per dose. In some embodiments, one, two, or three doses are administered to the subject. In some embodiments, one, two, or three doses are administered over a seven-day period. In some embodiments, 15 mL of the treatment product is suspended in 100 mL of saline and administered intravenously over a one-hour period. In some embodiments, 15 mL of the treatment product is suspended in 100 mL of saline and administered intravenously over one-hour periods on days 0, 2, and 4. In some embodiments, the administration is repeated one, two, three, four, or five times.

[0082] In some embodiments disclosed herein, treatment is provided for recipients of solid abdominal organ transplants who, despite routine immunosuppression, have evidence of graft-versus-host disease (GVHD) or solid abdominal graft rejection, and who are treated with a bone marrow MSC-derived EV therapeutic product (Direct Biologics, Austin TX). In some embodiments, the MSC-derived EVs disclosed herein are contained in readily storable, off-the-shelf products that are easy to administer and have a high safety profile. As described herein, MSC-derived EVs were successfully used to treat seven transplant patients with evidence of acute intestinal graft rejection and GVHD. No adverse or serious adverse events occurred, and all patients achieved complete clinical and histological recovery after delivery of the therapeutic product. The MSC-derived EV implementations disclosed herein represent a novel and safe therapeutic agent with the potential efficacy shown herein for patients with allogeneic graft rejection in the presence of infectious complications or GVHD. The implementations herein of extracellular vesicles derived from mesenchymal stem cells cultured under hypoxic, low-glucose, low-pH conditions provide an anti-inflammatory and immunomodulatory product for the transplant patient population.

[0083] definition As used herein and in the appended claims, unless otherwise expressly specified, the singular forms “a / an” and “the” include plural indicators. It should also be noted that, unless otherwise expressly specified, the term “or” is generally used in its meaning including “and / or”. The terms “and / or” and “any combination thereof” as used herein, and their grammatical equivalents, may be used interchangeably.

[0084] The term "about" or "approximately" can refer to an acceptable range of error for a particular value, which can depend in part on how the value was measured or determined, such as limitations of the measurement system. For example, "about" can refer to a standard deviation of less than or greater than 1. Alternatively, "about" can refer to a range of up to 20%, up to 10%, up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or methods, the term can refer to an order of magnitude, within 5 times, or within 2 times the value. When a particular value is described in the application and claims, unless otherwise stated, the term "about" should be presumed to refer to an acceptable range of error for the particular value.

[0085] As used in this specification and claims, the terms “comprising” (and any form of “comprising”, such as “comprise”, “comprises”), “having” (and any form of “having”, such as “having”, “suffering”), “including” (and any form of “including”, such as “includes”, “include”), or “containing” (and any form of “containing”, such as “contains”, “contain”) are inclusive or open-ended and do not exclude additional, unlisted elements or method steps.

[0086] The terms “individual,” “patient,” or “object” are used interchangeably. None of these terms require or are limited to situations characterized by supervision (e.g., continuous or intermittent) by a healthcare worker (e.g., physician, registered nurse, nurse practitioner, physician assistant, caregiver, or hospice worker).

[0087] Non-restrictive numbering implementation plan 1. A method for treating graft-versus-host disease in a subject, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition, said therapeutic mesenchymal stem cell (MSC) secretome composition being prepared by a method comprising: (a) Culture bone marrow-derived MSCs under the following conditions to produce MSC conditioned medium: (i) oxygen tension below 5%; and (ii) Culture media with a pH below 7; (b) Harvesting the MSC conditioned medium; and (c) Prepare the MSC conditioned medium to produce the therapeutic MSC secretome composition, wherein the therapeutic MSC secretome composition comprises proteins and extracellular vesicles produced by the bone marrow-derived MSCs in step (a).

[0088] 2. The method as described in implementation scheme 1, wherein the subject has received an organ transplant or allogeneic graft.

[0089] 3. The method as described in embodiment 2, wherein the subject has received a modified multi-organ allogeneic graft, a single intestinal allogeneic graft, or a single liver allogeneic graft.

[0090] 4. The method of any one of embodiments 1 to 3, wherein the subject exhibits one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection.

[0091] 5. The method as described in any one of embodiments 1 to 4, wherein the subject exhibits one or more signs or symptoms selected from the following: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, pruritus, spasms, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof.

[0092] 6. The method of any one of embodiments 1 to 5, wherein the subject is receiving corticosteroids.

[0093] 7. The method as described in embodiment 6, wherein the subject is receiving a high dose of corticosteroids.

[0094] 8. The method as described in any one of embodiments 1 to 7, wherein the subject experiences improvement in one or more of the following signs or symptoms after treatment: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, pruritus, spasms, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof.

[0095] 9. The method of any one of embodiments 1 to 8, wherein the dose of the therapeutic MSC secretory group composition administered to the subject is a cell equivalent dose of 700,000 to 7,000,000 cells / kg.

[0096] 10. The method of any one of embodiments 1 to 9, wherein the culture medium is serum-free.

[0097] 11. The method of any one of embodiments 1 to 10, wherein the culture medium has a glucose concentration of less than 4.5 g / L.

[0098] 12. The method of any one of embodiments 1 to 11, wherein at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63+CD9-CD81-.

[0099] 13. The method according to any one of embodiments 1 to 12, wherein the therapeutic MSC secretome composition comprises one or more of the following proteins: ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1, OPN, PAI-1, DAPP1, cathepsin B, brain signaling protein 6C, PDGF Rα, sorting protein, serine protease inhibitor B6, Dkk-3, coagulation regulatory protein, PF4, MIF, periosteal protein, furin, TIMP-1, gypsum proteoglycan, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, photoglucan, TIMP-2, SLITRK5, FAP, leptospirin, DPPII, cIAP-1, pentameric protein 3, lactone, enkephalin, albumin, galactolectin-1, UNC5H3, IL-20 Rβ, SREC-II, JAM-C, TNF. RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, convolutional protein A4, EMMPRIN, p53, brain signaling protein 7A, NKp80, cysteine ​​protease protein B, osteoadhesion, mid-term factor, calreticulin, osteoactivin, asparagine endopeptidase, TAZ, cathepsin L, RBP4, serine protease inhibitor A4, JAM-A, MCSF, LIMPII, OPG, IL-22, galactolectin-3, MOG, trypsin 3, SIRP α, cohesin-4, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, transmembrane peptidase A, IL-1 F10, bIG-H3, GPR115, TGFb1, liver glycoside A4, CD109, serine protease inhibitor F1, IGFBP-6, HS3ST4, aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, cathepsin B, IGFBP-2, brain signaling protein 6C, IGF-2, PDGFR α, sorting protein, serine protease inhibitor B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, coagulation regulatory protein, endosaccharides, IGFBP-3, RGM-C, PF4, MIF, TGM4, periosteal protein, furin, TIMP-1, PAPP-A, dermal proteoglycan, PCK1, arylsulfatase A, CD99, CA2, PRDX4, transferrin, DcR3, GP73, LAIR2, ULBP-4, optical proteoglycan, TIMP-2, TFPI, SOX2, SLITRK5, FAP, spinal cord protein, ENPP-2, CD97, CTACK, integrin α1, EXTL3, IL-18BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin I, IGF-2R, ADAMTS L1-1, Erythropoietin, Convolutional D1, DNMT3A, BCL-2, CL-P1, Hepatocyte Glycol-B3, FABP6, CHI3L1, FCRLS, TFF3, Nephlebotomycin, DPPII, cIAP-1, PDGF Rb, Pentanoic Acid 3, Angiotensinogen, Follicle-Stapressin, CF VII, Persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Lactone, Granulosin Progenitor, PCSK2, GKN1, IL-18, Enkephalin, Stabilin-2, IL-17 RD, albumin, follicle-stimulating protein 1, MMP-10, FKBP51, LRRC4, Pref-1, galactoglobin-1, troponin C, UNC5H3, FLRT2, CD314, brain signaling protein 6B, cytokinin-4, CD27 ligand, IL-20 Rβ, brain signaling protein 6A, TSK, cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, norin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Glycopropyl peptide, Activator Protein A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Phosphatidylinositol Proteoglycan 2, MSP R, DSCAM, Membrane-type Serine Protease, KIR2DL3, CD30, Salivary Agglutinin-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nestin-1, CD2, Kallikrein 1, TSLPR, LAMP1, TROY, VCAM-1, Salivary agglutinin-11, S100A1, PAR1, Thyroid peroxidase, Aminopeptidase P2, IL-1RI, ADAMS, OSM Rβ, Platelet-Reactive Protein-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granulase B, Lipocontin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Lipoprotein, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1α, OX40 ligand, Pax3, UCH-L3, cMASP3, Langerin, desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, conglomerate A4, brain signaling protein 4D, ROBO2, PDX-1, APRIL, neuronal rank protein, transmembrane protein-2 with ring structure, EMMPRIN, activating protein RIB, neuroligand 2, epithelial regulatory protein, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, extracellular matrix protein-2, sFRP-3, p53, EphB3, NCK1, brain signaling protein 7A, NKp80, prolactin, cysteine ​​agonist protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Brain signaling protein 6D, FGF-3, GATA-4, VAP-A, CHST2, Pregnancy-associated plasma protein-2, Cohesin-3, Serpentin-1, AKR1C4, Oligosin-2, Osteoadhesin, NKp44, Thyroglobulin, IL-21R, Chemokines, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin α 5. VAMP-2, FLRG, Ret metaphase factor, CD73, TRAP, proGRP, granzyme H, PRX2, p2'7, salivary lectin-6, dendritic cell-associated C-type lectin-1, CD51, Notch-1, calreticulin, DR3, DCTN1, CDC25B, bone activator, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, cathepsin V, testosterone-2, phosphatidylinositol proteoglycan 5, CD6, salivary lectin-2, asparagine endopeptidase, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 β, TAFA1, podocyte protein, RalA, CRELD2, GRAP2, SP-D, BID, GFR α-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, cathepsin L, 6-Ckine, MIS RII, kallikrein 5, TGM3, FCAR, contactin-2, CD83, IL-1R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, disaccharide proteoglycan, neurodevelopmental factor, ILT4, uPAR, Axl, WIF-1, IL-7 Rα, GPR56, CEACAM-3, MCEMP1, FABP2, plexin B3, MEPE, activator RIIA, ANG-2, Cochlin, presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, stalkin-1, GUSB, nestin-2, IL-17F, SR-AI, TAFA2, N-cadherin, IL-17B, IL-17RC, MIP-3b, cysteine ​​protease C, cysteine ​​protease D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, intestinal peptidase, cathepsin E, TSLP, TCN2, GDF-15, epidermal morphogenetics, GRKS, PD-1, serine protease inhibitor A4, ADAM23, NOV, galactoglobulin-2, neuronal surface protein 3β, TLR3, Sirtuin2, Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, lymphocyte chemokine, cysteine ​​protease protein SN, JAM-A, calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, kallikrein 14, extracellular matrix protein-3, Oligo2, kallikrein 12, CA13, IL-9, stalk protein-3, MPIF-1, cysteine ​​protease protein S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, transmembrane serine protease, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, brain signaling protein 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-1ra, Flatfoot protein, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenosynovin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Salivary lectin-9, ICAM-3, Cysteine ​​protease inhibitor SA, Galactohemagglutinin-4, Pepsinogen II, Desmosome core protein-3, Stalk protein-4, SCF, Serine protease inhibitor A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galactohemagglutinin-9, vWF-A2, TACE, Activator protein RIM, Cathepsin S, LDLR, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, phosphatidylinositol proteoglycan 1, LAP (TGFb1), Gash, contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, neural epithelial stem cell protein, TPO, ErbB3, Kirrel3, FLRT1, galactoglobulin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 Rα 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, desmosome core protein 2, agglutinin, synaptic fusion protein 4, VAMP-1, stalkin-2, FGF-21, Flt-3, GFAP, TIM-1, inhibin A, cadherin-4, PIGF-2, neurogranulin, HE4, IL-23 R, galactoglobulin-7, GALNT3, GITR L, CD14, R-vertebral protein 2, CK19, cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, salivary immunoglobulin-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-1a, PDGF-AB, GPVI, CD40, SCF R, platelet-reactive protein-5, IL-1 RII, neurofeedin-2, cadherin-13, E-selectin, GITR, WISP-1, renin, AgRP, MDL-1, ROBO3, RANTES, endothelial cell-specific molecules, granzyme, hCGb, mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, trypsin 3, ARSB, LIF Rα, BAFFR, CD157, granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP α, urinary modulating protein, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3, IL-32 α, TGFb3FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, fibrinogen-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Synaptic Fusion Protein 6, GRO, Bcl-w, Lipocrine-2, PDGF-AA, IL-2 Ra, Angiopoietin, LYVE-1, CD4, RAGE, CDNF, Short Proteoglycan, NAP-2, PU.1, EDAR, ADAMTS13, Kynurenase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, procalcitonin, SOX15, kallikrein 11, BCMA, dendritic cell-associated C-lectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, resistin, HVEM, ENPP-7, cohesin-4, IL-2 Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, brain signaling protein 4G, IL-12p40, total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, endothelial glycoprotein, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, galactolectin-8, CA4, cysteine ​​protease protein EM, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7 or IL-13.

[0100] 14. The method according to any one of embodiments 1 to 13, wherein the therapeutic MSC secretome composition comprises one or more of the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-mi R-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR- 132-3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-1 81a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19 a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-22 1. hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p,hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3 p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa -miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-3 61-5p, hsa-miR-376a-3p, hsa-miR-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424- 5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR -574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, h sa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p or hsa-miR-99b-5p. ,

[0101] 15. A method for treating graft-versus-host disease in a subject, the method comprising administering to the subject a composition comprising a therapeutic MSC secretome composition containing extracellular vesicles, wherein at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63+CD9-CD81-.

[0102] 16. The method of embodiment 15, wherein the therapeutic MSC secretome composition further comprises one or more of the following proteins: ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1, OPN, PAI-1, DAPP1, cathepsin B, brain signaling protein 6C, PDGF Rα, sorting protein, serine protease inhibitor B6, Dkk-3, coagulation regulatory protein, PF4, MIF, periosteal protein, furin, TIMP-1, tectonic proteoglycan, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, luciferin, TIMP-2, SLITRK5, FAP, leptospirin, DPPII, cIAP-1, pentameric protein 3, lactone, enkephalin, albumin, galactolectin-1, UNC5H3, IL-20 Rβ, SREC-II, JAM-C, TNF. RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, convolutional protein A4, EMMPRIN, p53, brain signaling protein 7A, NKp80, cysteine ​​protease protein B, osteoadhesion, mid-term factor, calreticulin, osteoactivin, asparagine endopeptidase, TAZ, cathepsin L, RBP4, serine protease inhibitor A4, JAM-A, MCSF, LIMPII, OPG, IL-22, galactolectin-3, MOG, trypsin 3, SIRP α, cohesin-4, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, transmembrane peptidase A, IL-1 F10, bIG-H3, GPR115, TGFb1, liver glycoside A4, CD109, serine protease inhibitor F1, IGFBP-6, HS3ST4, aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, cathepsin B, IGFBP-2, brain signaling protein 6C, IGF-2, PDGFR α, sorting protein, serine protease inhibitor B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, coagulation regulatory protein, endosaccharides, IGFBP-3, RGM-C, PF4, MIF, TGM4, periosteal protein, furin, TIMP-1, PAPP-A, dermal proteoglycan, PCK1, arylsulfatase A, CD99, CA2, PRDX4, transferrin, DcR3, GP73, LAIR2, ULBP-4, optical proteoglycan, TIMP-2, TFPI, SOX2, SLITRK5, FAP, spinal cord protein, ENPP-2, CD97, CTACK, integrin α1, EXTL3, IL-18BPa, PD-L2, PSMA, IL-20 Ra, Glyoxalase II, Trypsin I, IGF-2R, ADAMTS L1-1, Erythropoietin, Convolutional D1, DNMT3A, BCL-2, CL-P1, Hepatocyte Glycol-B3, FABP6, CHI3L1, FCRLS, TFF3, Nephlebotomycin, DPPII, cIAP-1, PDGF Rb, Pentanoic Acid 3, Angiotensinogen, Follicle-Stapressin, CF VII, Persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Lactone, Granulosin Progenitor, PCSK2, GKN1, IL-18, Enkephalin, Stabilin-2, IL-17 RD, albumin, follicle-stimulating protein 1, MMP-10, FKBP51, LRRC4, Pref-1, galactoglobin-1, troponin C, UNC5H3, FLRT2, CD314, brain signaling protein 6B, cytokinin-4, CD27 ligand, IL-20 Rβ, brain signaling protein 6A, TSK, cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, norin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Glycopropyl peptide, Activator Protein A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Phosphatidylinositol Proteoglycan 2, MSP R, DSCAM, Membrane-type Serine Protease, KIR2DL3, CD30, Salivary Agglutinin-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nestin-1, CD2, Kallikrein 1, TSLPR, LAMP1, TROY, VCAM-1, Salivary agglutinin-11, S100A1, PAR1, Thyroid peroxidase, Aminopeptidase P2, IL-1RI, ADAMS, OSM Rβ, Platelet-Reactive Protein-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granulase B, Lipocontin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, Lipoprotein, AIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1α, OX40 ligand, Pax3, UCH-L3, cMASP3, Langerin, desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, conglomerate A4, brain signaling protein 4D, ROBO2, PDX-1, APRIL, neuronal rank protein, transmembrane protein-2 with ring structure, EMMPRIN, activating protein RIB, neuroligand 2, epithelial regulatory protein, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, extracellular matrix protein-2, sFRP-3, p53, EphB3, NCK1, brain signaling protein 7A, NKp80, prolactin, cysteine ​​agonist protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Brain signaling protein 6D, FGF-3, GATA-4, VAP-A, CHST2, Pregnancy-associated plasma protein-2, Cohesin-3, Serpentin-1, AKR1C4, Oligosin-2, Osteoadhesin, NKp44, Thyroglobulin, IL-21R, Chemokines, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin α 5. VAMP-2, FLRG, Ret metaphase factor, CD73, TRAP, proGRP, granzyme H, PRX2, p2'7, salivary lectin-6, dendritic cell-associated C-type lectin-1, CD51, Notch-1, calreticulin, DR3, DCTN1, CDC25B, bone activator, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, cathepsin V, testosterone-2, phosphatidylinositol proteoglycan 5, CD6, salivary lectin-2, asparagine endopeptidase, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 β, TAFA1, podocyte protein, RalA, CRELD2, GRAP2, SP-D, BID, GFR α-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, cathepsin L, 6-Ckine, MIS RII, kallikrein 5, TGM3, FCAR, contactin-2, CD83, IL-1R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, disaccharide proteoglycan, neurodevelopmental factor, ILT4, uPAR, Axl, WIF-1, IL-7 Rα, GPR56, CEACAM-3, MCEMP1, FABP2, plexin B3, MEPE, activator RIIA, ANG-2, Cochlin, presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, stalkin-1, GUSB, nestin-2, IL-17F, SR-AI, TAFA2, N-cadherin, IL-17B, IL-17RC, MIP-3b, cysteine ​​protease C, cysteine ​​protease D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, intestinal peptidase, cathepsin E, TSLP, TCN2, GDF-15, epidermal morphogenetics, GRKS, PD-1, serine protease inhibitor A4, ADAM23, NOV, galactoglobulin-2, neuronal surface protein 3β, TLR3, Sirtuin2, Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, lymphocyte chemokine, cysteine ​​protease protein SN, JAM-A, calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, kallikrein 14, extracellular matrix protein-3, Oligo2, kallikrein 12, CA13, IL-9, stalk protein-3, MPIF-1, cysteine ​​protease protein S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, transmembrane serine protease, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, brain signaling protein 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-1ra, Flatfoot protein, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenosynovin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Salivary lectin-9, ICAM-3, Cysteine ​​protease inhibitor SA, Galactohemagglutinin-4, Pepsinogen II, Desmosome core protein-3, Stalk protein-4, SCF, Serine protease inhibitor A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galactohemagglutinin-9, vWF-A2, TACE, Activator protein RIM, Cathepsin S, LDLR, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, phosphatidylinositol proteoglycan 1, LAP (TGFb1), Gash, contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, neural epithelial stem cell protein, TPO, ErbB3, Kirrel3, FLRT1, galactoglobulin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 Rα 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, desmosome core protein 2, agglutinin, synaptic fusion protein 4, VAMP-1, stalkin-2, FGF-21, Flt-3, GFAP, TIM-1, inhibin A, cadherin-4, PIGF-2, neurogranulin, HE4, IL-23 R, galactoglobulin-7, GALNT3, GITR L, CD14, R-vertebral protein 2, CK19, cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, salivary immunoglobulin-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-1a, PDGF-AB, GPVI, CD40, SCF R, platelet-reactive protein-5, IL-1 RII, neurofeedin-2, cadherin-13, E-selectin, GITR, WISP-1, renin, AgRP, MDL-1, ROBO3, RANTES, endothelial cell-specific molecules, granzyme, hCGb, mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, trypsin 3, ARSB, LIF Rα, BAFFR, CD157, granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP α, urinary modulating protein, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3, IL-32 α, TGFb3FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, fibrinogen-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Synaptic Fusion Protein 6, GRO, Bcl-w, Lipocrine-2, PDGF-AA, IL-2 Ra, Angiopoietin, LYVE-1, CD4, RAGE, CDNF, Short Proteoglycan, NAP-2, PU.1, EDAR, ADAMTS13, Kynurenase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, procalcitonin, SOX15, kallikrein 11, BCMA, dendritic cell-associated C-lectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, resistin, HVEM, ENPP-7, cohesin-4, IL-2 Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, brain signaling protein 4G, IL-12p40, total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, endothelial glycoprotein, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, galactolectin-8, CA4, cysteine ​​protease protein EM, FUT8, B7-H3, GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7 or IL-13.

[0103] 17. The method according to embodiment 15 or 16, wherein the extracellular vesicles comprise one or more of the following nucleic acids: hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hsa-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir-10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-miR-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p, hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1’7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19’7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25,hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR -29a-3p, hsa-miR-29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c -5p, hsa-mir-30d, hsa-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR -31-5p, hsa-miR-320a, hsa-miR-342-3p, hsa-miR-345-5p, hsa-miR-34a-5p, hsa-miR-361-5p , hsa-miR-376a-3p, hsa-miR-376c-3p, hsa-miR-423-3p, hsa-miR-423-5p, hsa-miR-424-5p, h sa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-57 4-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa -miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p or hsa-miR-99b-5p. ,

[0104] 18. The method of any one of embodiments 15 to 17, wherein the subject has received an organ transplant or allogeneic graft.

[0105] 19. The method of embodiment 18, wherein the subject has received a modified multi-organ allogeneic graft, a single intestinal allogeneic graft, or a single liver allogeneic graft.

[0106] 20. The method of any one of embodiments 15 to 19, wherein the subject exhibits one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection.

[0107] 21. The method as described in any one of embodiments 15 to 20, wherein the subject exhibits one or more signs or symptoms selected from the following: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, pruritus, spasms, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof.

[0108] 22. The method of any one of embodiments 15 to 21, wherein the subject is receiving corticosteroids.

[0109] 23. The method of embodiment 22, wherein the subject is receiving a high dose of corticosteroids.

[0110] 24. The method as described in any one of embodiments 15 to 23, wherein the subject experiences improvement in one or more of the following signs or symptoms after treatment: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, pruritus, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof.

[0111] 25. The method as described in any one of embodiments 15 to 24, wherein the subject shows improvement within 24 hours of application of the treatment product.

[0112] 26. A method for treating graft-versus-host disease in a subject, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles (EVs), wherein the therapeutic MSC secretome comprises: (i) ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1, OPN, PAI-1, DAPP1, cathepsin B, brain signaling protein 6C, PDGF R α, Sorting protein, serine protease inhibitor B6, Dkk-3, coagulation regulator protein, PF4, MIF, periosteal protein, furin, TIMP-1, dermal proteoglycan, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, photoglycan, TIMP-2, SLITRK5, FAP, leptospirin, DPPII, cIAP-1, pentameric protein 3, lactone, enkephalin, albumin, galactoglobin-1, UNC5H3, IL-20, β-reactive protein, SREC-II, JAM-C, TNF-α, htPAPP-A, eNOS, MSP, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, convolutional protein A4, EMMPRIN, p53, brain signaling protein 7A, NKp80, cysteine ​​protease protein B, osteoadhesion, mid-term factor, calreticulin, osteoactivin, asparagine endopeptidase, TAZ, cathepsin L, RBP4, serine protease inhibitor A4, JAM-A, MCSF, LIMPII, OPG, IL-22, galactoglobulin-3, MOG, trypsin 3, SIRP α, cohesin-4, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, transmembrane peptidase A, IL-1 F10, bIG-H3, GPR115, TGFb1, liver glycoside A4, CD109, serine protease inhibitor F1, IGFBP-6, HS3ST4, aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, cathepsin B, IGFBP-2, brain signaling protein 6C, IGF-2, PDGFR α, sorting protein, serine protease inhibitor B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, coagulation regulatory protein, endosaccharides, IGFBP-3, RGM-C, PF4, MIF, TGM4, periosteal protein, furin protease, TIMP-1, PAPP-A, dermal proteoglycans, PCK1, arylsulfatase A, CD99, CA2, PRDX4, transferrin, DcR3, GP73, LAIR2, ULBP-4, optical proteoglycans, TIMP-2, TFPI, SOX2, SLITRK5, FAP, spinal proteinENPP-2, CD97, CTACK, integrin α1, EXTL3, IL-18, BPa, PD-L2, PSMA, IL-20 Ra, glyoxalase II, trypsin I, IGF-2R, ADAMTSL-1, erythropoietin, plexin D1, DNMT3A, BCL-2, CL-P1, hepatocyte glycoside-B3, FABP6, CHI3L1, FCRLS, TFF3, leptospirin, DPPII, cIAP-1, PDGF Rb, pentameric protein 3, angiotensinogen, follicle-stimulating hormone, CF VII, persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, endothelin, granulosin precursor, PCSK2, GKN1, IL-18, enkephalin, stabilizer-2, IL-17 RD, albumin, follicle-stimulating protein 1, MMP-10, FKBP51, LRRC4, Pref-1, galactoglobin-1, troponin C, UNC5H3, FLRT2, CD314, brain signaling protein 6B, cytokinin-4, CD27 ligand, IL-20 Rβ, brain signaling protein 6A, TSK, cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, norin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Glycopropyl peptide, Activator Protein A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Phosphatidylinositol Proteoglycan 2, MSP R, DSCAM, Membrane-type Serine Protease, KIR2DL3, CD30, Salivary Agglutinin-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nestin-1, CD2, Kallikrein 1, TSLPR, LAMP1, TROY, VCAM-1, Salivary agglutinin-11, S100A1, PAR1, Thyroid peroxidase, Aminopeptidase P2, IL-1RI, ADAMS, OSM Rβ, Platelet-Reactive Protein-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granulase B, Lipocontin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, LipoproteinAIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 α, OX40 ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Convolutional protein A4, Brain signaling protein 4D, ROBO2, PDX-1, APRIL, Neural rank protein, cyclic transmembrane protein-2, EMMPRIN, activator protein RIB, neuroligand 2, epithelial regulatory protein, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Extracellular matrix protein-2, sFRP-3, p53, EphB3, NCK1, Brain signaling protein 7A, NKp80, Prolactin, Cysteine ​​protease protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Brain signaling protein 6D, FGF-3, GATA-4, VAP-A, CHST2, Pregnancy-associated plasma protein-2, Cohesin-3, Serpentin-1, AKR1C4, Oligosin-2, Osteoadhesin, NKp44, Thyroglobulin, IL-21R, Chemokines, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin α 5. VAMP-2, FLRG, Ret metaphase factor, CD73, TRAP, proGRP, granzyme H, PRX2, p2'7, salivary lectin-6, dendritic cell-associated C-type lectin-1, CD51, Notch-1, calreticulin, DR3, DCTN1, CDC25B, bone activator, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, cathepsin V, testosterone-2, phosphatidylinositol proteoglycan 5, CD6, salivary lectin-2, asparagine endopeptidase, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 β, TAFA1, podocyte protein, RalA, CRELD2, GRAP2, SP-D, BID, GFR α-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, cathepsin L, 6-Ckine, MIS RII, kallikrein 5, TGM3, FCAR, contactin-2, CD83, IL-1R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, disaccharide proteoglycan, neurodevelopmental factor, ILT4, uPAR, Axl, WIF-1IL-7 Rα, GPR56, CEACAM-3, MCEMP1, FABP2, plexin B3, MEPE, activator RIIA, ANG-2, Cochlin, progerin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, stalkin-1, GUSB, nestin-2, IL-17F, SR-AI, TAFA2, N-cadherin, IL-17B, IL-17RC, MIP-3b, cysteine ​​protease C, cysteine ​​protease D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, intestinal peptidase, cathepsin E, TSLP, TCN2, GDF-15, epidermal morphogenetics, GRKS, PD-1, serine protease inhibitor A4, ADAM23, NOV, galactoglobulin-2, neuronal surface protein 3β, TLR3, Sirtuin2, Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, lymphocyte chemokine, cysteine ​​protease protein SN, JAM-A, calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, kallikrein 14, extracellular matrix protein-3, Oligo2, kallikrein 12, CA13, IL-9, stalk protein-3, MPIF-1, cysteine ​​protease protein S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, transmembrane serine protease, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, brain signaling protein 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-1ra, Flatfoot protein, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenosynovin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Salivary lectin-9, ICAM-3, Cysteine ​​protease inhibitor SA, Galactohemagglutinin-4, Pepsinogen II, Desmosome core protein-3, Stalk protein-4, SCF, Serine protease inhibitor A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galactohemagglutinin-9, vWF-A2, TACE, Activator protein RIM, Cathepsin S, LDL RBMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, phosphatidylinositol proteoglycan 1, LAP (TGFb1), Gash, contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, neural epithelial stem cell protein, TPO, ErbB3, Kirrel3, FLRT1, galactoglobulin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 Rα 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, desmosome core protein 2, agglutinin, synaptic fusion protein 4, VAMP-1, stalkin-2, FGF-21, Flt-3, GFAP, TIM-1, inhibin A, cadherin-4, PIGF-2, neurogranulin, HE4, IL-23 R, galactoglobulin-7, GALNT3, GITR L, CD14, R-vertebral protein 2, CK19, cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, salivary immunoglobulin-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-1a, PDGF-AB, GPVI, CD40, SCF R, platelet-reactive protein-5, IL-1 RII, neurofeedin-2, cadherin-13, E-selectin, GITR, WISP-1, renin, AgRP, MDL-1, ROBO3, RANTES, endothelial cell-specific molecules, granzyme, hCGb, mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, trypsin 3, ARSB, LIF Rα, BAFFR, CD157, granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP α, urinary modulatory proteins, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3, IL-32 α, TGFb3, FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, fibrinogen-1, Reg4ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Synaptic Fusion Protein 6, GRO, Bcl-w, Lipocrine-2, PDGF-AA, IL-2 Ra, Angiopoietin, LYVE-1, CD4, RAGE, CDNF, Short Proteoglycan, NAP-2, PU.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dendritic Cell-Associated C-Lectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Cohesin-Glycan-4, IL-2 Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, brain signaling protein 4G, IL-12p40, total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, endothelial glycoprotein, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, galactolectin-8, CA4, cysteine ​​protease protein EM, FUT8, B7-H 3. GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7 or IL-13 or a combination of two or more thereof; (ii) hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hs a-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir- 10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-mi R-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p , hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p,hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-mi R-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa- mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p hsa-miR-320a hsa-miR-342-3p hsa-miR-345-5p hsa-miR-34a-5p hsa-miR-361-5p hsa-miR-376a-3p hsa-miR-376c-3p hsa-miR-423-3p hsa -miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663ahsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p or hsa-miR-99b-5p, or a combination of two or more thereof; or both (iii), (i) and (ii).

[0113] 27. A method for treating graft-versus-host disease, the method comprising administering a therapeutic MSC secretome composition to the subject.

[0114] 28. A method for treating allogeneic graft rejection, the method comprising administering a therapeutic MSC secretome composition to the subject.

[0115] 29. A method for alleviating symptoms associated with graft-versus-host disease in a subject, the method comprising administering a therapeutic MSC secretome composition to the subject.

[0116] 30. A method for improving graft-versus-host disease and / or allogeneic graft rejection of one or more signs or symptoms selected from the following: crypt cell apoptosis, crypt loss, ulceration, allogeneic graft cell rejection, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, pruritus, spasms, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof.

[0117] 31. The method of any one of embodiments 27-30, wherein the extracellular vesicles in the therapeutic MSC secretome composition are CD63+CD9-CD81-.

[0118] 32. The method of any one of embodiments 27-31, wherein the therapeutic MSC secretory group comprises (i) ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1, OPN, PAI-1, DAPP1, cathepsin B, brain signaling protein 6C, PDGF Rα, sorting protein, serine protease inhibitor B6, Dkk-3, coagulation regulator protein, PF4, MIF, periosteal protein, furin, TIMP-1, gypsum proteoglycan, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, photoglucan, TIMP-2, SLITRK5, FAP, leptospirin, DPPII, cIAP-1, pentameric protein 3, lactone, enkephalin, albumin, galactolectin-1, UNC5H3, IL-20 Rβ, SREC-II, JAM-C, TNF RI, htPAPP-A, eNOS, MSP R, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, convolutional protein A4, EMMPRIN, p53, brain signaling protein 7A, NKp80, cysteine ​​protease protein B, osteoadhesion, mid-term factor, calreticulin, osteoactivin, asparagine endopeptidase, TAZ, cathepsin L, RBP4, serine protease inhibitor A4, JAM-A, MCSF, LIMPII, OPG, IL-22, galactolectin-3, MOG, trypsin 3, SIRP α, cohesin-4, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, transmembrane peptidase A, IL-1 F10, bIG-H3, GPR115, TGFb1, liver glycoside A4, CD109, serine protease inhibitor F1, IGFBP-6, HS3ST4, aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, cathepsin B, IGFBP-2, brain signaling protein 6C, IGF-2, PDGF Rα, sorting protein, serine protease inhibitor B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, coagulation regulatory protein, endosaccharides, IGFBP-3, RGM-C, PF4, MIF, TGM4, periostealin, furin, TIMP-1, PAPP-A, dermal proteoglycans, PCK1, arylsulfatase A, CD99, CA2, PRDX4, transferrin, DCR3, GP73, LAIR2, ULBP-4, luminous proteoglycans, TIMP-2, TFPI, SOX2, SLITRK5, FAP, spinal cord protein, ENPP-2, CD97, CTACK, integrin α1, EXTL3, IL-18, BPa, PD-L2, PSMAIL-20 Ra, Glyoxalase II, Trypsin I, IGF-2R, ADAMTS L1-1, Erythropoietin, Convolutional D1, DNMT3A, BCL-2, CL-P1, Hepatocyte Glycol-B3, FABP6, CHI3L1, FCRLS, TFF3, Sphingomyelin, DPPII, cIAP-1, PDGF Rb, Pentanoic Acid 3, Angiotensinogen, Follicle-Stapressin, CF VII, Persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, Lactone, Granulosin Progenitor, PCSK2, GKN1, IL-18, Enkephalin, Stabilin-2, IL-17 RD, albumin, follicle-stimulating protein 1, MMP-10, FKBP51, LRRC4, Pref-1, galactoglobin-1, troponin C, UNC5H3, FLRT2, CD314, brain signaling protein 6B, cytokinin-4, CD27 ligand, IL-20 Rβ, brain signaling protein 6A, TSK, cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, norin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Glycopropyl peptide, Activator Protein A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Phosphatidylinositol Proteoglycan 2, MSP R, DSCAM, Membrane-type Serine Protease, KIR2DL3, CD30, Salivary Agglutinin-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nestin-1, CD2, Kallikrein 1, TSLP R, LAMP1, TROY, VCAM-1, salivary immunoglobulin-11, S100A1, PAR1, thyroid peroxidase, aminopeptidase P2, IL-1 RI, ADAMS, OSM R β, platelet-reactive protein-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), granzyme B, lipoconazole, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, lipoprotein, AIF, SorCS2, SULT2A1, CD39L2, insulin R, HIF-1 α, OX40 ligandPax3, UCH-L3, cMASP3, Langerin, desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, conglomerate A4, brain signaling protein 4D, ROBO2, PDX-1, APRIL, neuronal rankin, transmembrane protein-2 with ring structure, EMMPRIN, activator protein RIB, neuroligand 2, epithelial regulatory protein, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, extracellular matrix protein-2, sFRP-3, p53, EphB3, NCK1, brain signaling protein 7A, NKp80, prolactin, cysteine ​​agonist protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Brain signaling protein 6D, FGF-3, GATA-4, VAP-A, CHST2, Pregnancy-associated plasma protein-2, Cohesin-3, Serpentin-1, AKR1C4, Oligosin-2, Osteoadhesin, NKp44, Thyroglobulin, IL-21R, Chemokines, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin α 5. VAMP-2, FLRG, Ret metaphase factor, CD73, TRAP, proGRP, granzyme H, PRX2, p27, salivary lectin-6, dendritic cell-associated C-type lectin-1, CD51, Notch-1, calreticulin, DR3, DCTN1, CDC25B, bone activator, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, cathepsin V, testosterone-2, phosphatidylinositol proteoglycan 5, CD6, salivary lectin-2, asparagine endopeptidase, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 β, TAFA1, podocyte protein, RalA, CRELD2, GRAP2, SP-D, BID, GFR α-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, cathepsin L, 6-Ckine, MIS RII, kallikrein 5, TGM3, FCAR, contactin-2, CD83, IL-1 R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, disaccharide proteoglycan, neurodevelopmental factor, ILT4, uPAR, Axl, WIF-1, IL-7 Rα, GPR56, CEACAM-3, MCEMP1, FABP2, conglomerate B3, MEPEActivating proteins RIIA, ANG-2, Cochlin, presenilin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, stalkin-1, GUSB, nestin-2, IL-17F, SR-AI, TAFA2, N-cadherin, IL-17B, IL-17RC, MIP-3b, cysteine ​​absorptase C, cysteine ​​absorptase D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, intestinal peptidase, cathepsin E, TSLP, TCN2, GDF-15, epidermal morphogenetics, GRKS, PD-1, serine protease inhibitor A4, ADAM23, NOV, galactolectin-2, neuronal surface protein 3β, TLR3, Sirtuin 2, Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, lymphocyte chemokines, cysteine ​​protease protein SN, JAM-A, calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, kallikrein 14, extracellular matrix protein-3, Olig2, kallikrein 12, CA13, IL-9, stalkin-3, MPIF-1, cysteine ​​protease protein S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, transmembrane serine protease, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, Brain signaling protein 4C, SorCS3, Tie-1, IL-31RA, Arginase 1, POGLUT1, IL-1ra, Flatfoot protein, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenosynovin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Salivary lectin-9, ICAM-3, Cysteine ​​protease inhibitor SA, Galactohemagglutinin-4, Pepsinogen II, Desmosome core protein-3, Stalk protein-4, SCF, Serine protease inhibitor A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, galactoglobulin-9, vWF-A2, TACE, activator protein RIM, cathepsin S, LDLR, BMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, phosphatidylinositol proteoglycan 1, LAP (TGFb1), Gash, contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10, Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, neural epithelial stem cell protein, TPO, ErbB3, Kirrel3, FLRT1, galactolectin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGFR2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22, Rα1, AMIGO2, HCC-1, TFPI-2, ULBP-2, desmosome core protein 2, agglutinin, synaptic fusion protein 4, VAMP-1, stalkin-2, FGF-21, Flt-3, GFAP, TIM-1, inhibin A, cadherin-4, PIGF-2, neurogranulin, HE4, IL-23, R, galactolectin-7, GALNT3, GITR L, CD14, R-vertebral protein 2, CK19, cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, salivary lectin-7, CD155, VEGF-C, TNF-α, PGRP-S, SDF-1α, PDGF-AB, GPVI, CD40, SCFR, platelet-reactive protein-5, IL-1 RII, neurofeltin-2, cadherin-13, E-selectin, GITR, WISP-1, renin, AgRP, MDL-1, ROBO3, RANTES, endothelial cell-specific molecules, granzyme, hCGb, mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, trypsin 3, ARSB, LIF Rα, BAFF R, CD157, granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP α, urinary modulatory proteins, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3, IL-32 α, TGFb3 FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, fibrin-1, Reg4, ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, insulin, synaptic fusion protein 6GRO, Bcl-w, lipocalin-2, PDGF-AA, IL-2 Ra, angiopoietin, LYVE-1, CD4, RAGE, CDNF, short proteoglycan, NAP-2, PU.1, EDAR, ADAMTS13, kynurenase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, procalcitonin, SOX15, kallikrein 11, BCMA, dendritic cell-associated C-lectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, resistin, HVEM, ENPP-7, cohesin-glycan-4, IL-2 Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, brain signaling protein 4G, IL-12p40, total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, endothelial glycoprotein, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, galactolectin-8, CA4, cysteine ​​protease protein EM, FUT8, B7-H 3. GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7 or IL-13 or a combination of two or more thereof; (ii) hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hs a-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir- 10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-mi R-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p , hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p, hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p,hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR- 203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa-mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR- 27a-3p、hsa-mir-27b、hsa-miR-27b-3p、hsa-miR-29a-3p、hsa-miR-29c-3 p、hsa-miR-30a-5p、hsa-miR-30a-5p、hsa-miR-30b-5p、hsa-miR-30c-5p、 hsa-mir-30d、hsa-miR-30d-5p、hsa-mir-30e、hsa-miR-30e-5p、hsa-miR- 31-3p、hsa-miR-31-5p、hsa-miR-320a、hsa-miR-342-3p、hsa-miR-345-5p hsa-miR-34a-5p hsa-miR-361-5p hsa-miR-376a-3p hsa-miR-376c-3p hsa-miR-423-3p hsa-miR-423-5p hsa-miR-424-5p hsa-miR-484 hsa- mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663a, hsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p or hsa-miR-99b-5p, or a combination of two or more thereof; or both (iii), (i) and (ii).

[0119] 33. The method of any one of embodiments 27-32, wherein the therapeutic MSC secretome composition is prepared by a method comprising the steps of: (a) culturing bone marrow-derived MSCs under conditions of: (i) oxygen tension below 5%; and (ii) a medium with a pH below 7; (b) harvesting the MSC conditioned medium; and (c) formulating the MSC conditioned medium to produce the therapeutic MSC secretome composition, wherein the therapeutic MSC secretome composition comprises proteins and extracellular vesicles produced by the bone marrow-derived MSCs in step (a).

[0120] 34. The method of any one of embodiments 27-33, wherein the subject has received an organ transplant or allogeneic graft.

[0121] 35. The method of embodiment 34, wherein the subject has received a modified multi-organ allogeneic graft, a single intestinal allogeneic graft, or a single liver allogeneic graft.

[0122] 36. The method of any one of embodiments 27-35, wherein the subject exhibits one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection.

[0123] 37. The method of any one of embodiments 27-36, wherein the subject exhibits one or more signs or symptoms selected from the following: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, pruritus, spasms, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof.

[0124] 38. The method of any one of embodiments 1-37, wherein the composition comprises 15 mL of the therapeutic MSC secretion group composition.

[0125] 39. The method of embodiment 38, wherein the composition is administered intravenously.

[0126] 40. A method for treating graft-versus-host disease in a subject, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles (EVs), wherein the composition comprises 15 mL of the therapeutic MSC secretome composition.

[0127] 41. A method for treating graft-versus-host disease in a subject, the method comprising administering to the subject a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition of extracellular vesicles (EVs), wherein the administration improves one or more signs or symptoms selected from: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, cramps, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof.

[0128] 42. The method of any one of embodiments 1-41, wherein the application improves inflammation.

[0129] 43. Use of a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles in graft-versus-host disease of a therapeutic subject, wherein at least 80% of the extracellular vesicles in the therapeutic MSC secretome composition are CD63+CD9-CD81-.

[0130] 44. Use of a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles (EVs) for the treatment of graft-versus-host disease in subjects of need, wherein the therapeutic mesenchymal stem cell (MSC) secretome composition comprises: (i) ferritin, NUP85, LAMP2, GPR115, serine protease inhibitor F1, OPN, PAI-1, DAPP1, cathepsin B, brain signaling protein 6C, PDGF R α, Sorting protein, serine protease inhibitor B6, Dkk-3, coagulation regulator protein, PF4, MIF, periosteal protein, furin, TIMP-1, dermal proteoglycan, PCK1, CD99, CD63, CD9, CD81, transferrin, DcR3, photoglycan, TIMP-2, SLITRK5, FAP, leptospirin, DPPII, cIAP-1, pentameric protein 3, lactone, enkephalin, albumin, galactoglobin-1, UNC5H3, IL-20, β-reactive protein, SREC-II, JAM-C, TNF-α, htPAPP-A, eNOS, MSP, TPP1, LAMP1, B2M, NCAM-1, HIF-1 α, ST6GAL1, CD99-L2, convolutional protein A4, EMMPRIN, p53, brain signaling protein 7A, NKp80, cysteine ​​protease protein B, osteoadhesion, mid-term factor, calreticulin, osteoactivin, asparagine endopeptidase, TAZ, cathepsin L, RBP4, serine protease inhibitor A4, JAM-A, MCSF, LIMPII, OPG, IL-22, galactoglobulin-3, MOG, trypsin 3, SIRP α, cohesin-4, IGFBP-4, IL-1 R6, GSTM1, NUP85, LAMP2, transmembrane peptidase A, IL-1 F10, bIG-H3, GPR115, TGFb1, liver glycoside A4, CD109, serine protease inhibitor F1, IGFBP-6, HS3ST4, aminopeptidase LRAP, OPN, PAI-1, DAPP1, GDF-9, cathepsin B, IGFBP-2, brain signaling protein 6C, IGF-2, PDGFR α, sorting protein, serine protease inhibitor B6, Dkk-3, CNTF, TSP-1, GM-CSF Ra, coagulation regulatory protein, endosaccharides, IGFBP-3, RGM-C, PF4, MIF, TGM4, periosteal protein, furin protease, TIMP-1, PAPP-A, dermal proteoglycans, PCK1, arylsulfatase A, CD99, CA2, PRDX4, transferrin, DcR3, GP73, LAIR2, ULBP-4, optical proteoglycans, TIMP-2, TFPI, SOX2, SLITRK5, FAP, spinal proteinENPP-2, CD97, CTACK, integrin α1, EXTL3, IL-18, BPa, PD-L2, PSMA, IL-20 Ra, glyoxalase II, trypsin I, IGF-2R, ADAMTSL-1, erythropoietin, plexin D1, DNMT3A, BCL-2, CL-P1, hepatocyte glycoside-B3, FABP6, CHI3L1, FCRLS, TFF3, leptospirin, DPPII, cIAP-1, PDGF Rb, pentameric protein 3, angiotensinogen, follicle-stimulating hormone, CF VII, persephin, TRAIL R1, THAP11, CD200, CLEC-2, AMIGO, IGFBP-5, PON1, SOX7, GALNT10, endothelin, granulosin precursor, PCSK2, GKN1, IL-18, enkephalin, stabilizer-2, IL-17 RD, albumin, follicle-stimulating protein 1, MMP-10, FKBP51, LRRC4, Pref-1, galactoglobin-1, troponin C, UNC5H3, FLRT2, CD314, brain signaling protein 6B, cytokinin-4, CD27 ligand, IL-20 Rβ, brain signaling protein 6A, TSK, cytokeratin-8, CHST3, Mcl-1, DPPIV, SREC-II, norin, JAM-C, Bcl-10, Wnt-4, LSECtin, Kell, TNF RI, PTP1B, htPAPP-A, IDO, PDGF-CC, Glycopropyl peptide, Activator Protein A, TLR2, SCCA2, FABP1, eNOS, SHP-1, ICOS, C1qTNF9, MMP-1, TC-PTP, IL-24, gp130, C-myc, LILRB4, BMP-2, MIA, CD34, CD63, CD9, CD81, IFNab R2, Phosphatidylinositol Proteoglycan 2, MSP R, DSCAM, Membrane-type Serine Protease, KIR2DL3, CD30, Salivary Agglutinin-10, CLEC-1, TPP1, Ubiquitin+1, ANGPTL4, TWEAK R, Nestin-1, CD2, Kallikrein 1, TSLPR, LAMP1, TROY, VCAM-1, Salivary agglutinin-11, S100A1, PAR1, Thyroid peroxidase, Aminopeptidase P2, IL-1RI, ADAMS, OSM Rβ, Platelet-Reactive Protein-2, SMPD1, B2M, MFRP, LRP-6, ST3GAL1, NCAM-1 (CD56), Granulase B, Lipocontin, IL-22BP, TPST2, PD-ECGF, LH, LEDGF, Cyr61, ULBP-3, IFNb, THSD1, FGF-23, LAMA4, LipoproteinAIF, SorCS2, SULT2A1, CD39L2, Insulin R, HIF-1 α, OX40 ligand, Pax3, UCH-L3, cMASP3, Langerin, Desmin, SOX9, ST6GAL1, MEP1B, CD99-L2, Convolutional protein A4, Brain signaling protein 4D, ROBO2, PDX-1, APRIL, Neural rank protein, cyclic transmembrane protein-2, EMMPRIN, activator protein RIB, neuroligand 2, epithelial regulatory protein, CASA, MMP-12, GALNT2, CEACAM-5, VEGF R1, DSPG3, SorCS1, Extracellular matrix protein-2, sFRP-3, p53, EphB3, NCK1, Brain signaling protein 7A, NKp80, Prolactin, Cysteine ​​protease protein B, Sirtuin 1, FGF-16, FGF R5, NQO-1, Brain signaling protein 6D, FGF-3, GATA-4, VAP-A, CHST2, Pregnancy-associated plasma protein-2, Cohesin-3, Serpentin-1, AKR1C4, Oligosin-2, Osteoadhesin, NKp44, Thyroglobulin, IL-21R, Chemokines, EphA1, CD48, MICB, FGF-5, TRANCE, CES2, ULBP-1, Integrin α 5. VAMP-2, FLRG, Ret metaphase factor, CD73, TRAP, proGRP, granzyme H, PRX2, p2'7, salivary lectin-6, dendritic cell-associated C-type lectin-1, CD51, Notch-1, calreticulin, DR3, DCTN1, CDC25B, bone activator, ACE, CA125, HAO-1, ​​PSMA1, FCRLB, BMP-9, CRIM1, LIF, SPINK1, EphB6, RGM-B, HS3ST1, ROR1, CMG-2, 4-1BB ligand, L1CAM-2, p63, cathepsin V, testosterone-2, phosphatidylinositol proteoglycan 5, CD6, salivary lectin-2, asparagine endopeptidase, PRELP, CES1, TAZ, NSE, TECK, HTRA2, HIF-1 β, TAFA1, podocyte protein, RalA, CRELD2, GRAP2, SP-D, BID, GFR α-2, Notch-3, VEGF R3, DLL4, TGFb2, LIGHT, XIAP, ST8SIA1, cathepsin L, 6-Ckine, MIS RII, kallikrein 5, TGM3, FCAR, contactin-2, CD83, IL-1R3, SALM4, GBA3, ROBO4, OSCAR, VEGF, IGSF3, disaccharide proteoglycan, neurodevelopmental factor, ILT4, uPAR, Axl, WIF-1IL-7 Rα, GPR56, CEACAM-3, MCEMP1, FABP2, plexin B3, MEPE, activator RIIA, ANG-2, Cochlin, progerin 1, NPTXR, SLAM, COMT, SPHK1, RBP4, stalkin-1, GUSB, nestin-2, IL-17F, SR-AI, TAFA2, N-cadherin, IL-17B, IL-17RC, MIP-3b, cysteine ​​protease C, cysteine ​​protease D, AMSH, FcERI, CLEC10A, HGF R, ANG-1, prolactin R, FGF-20, CD28, Nogo-A, HSD17B1, IL-19, intestinal peptidase, cathepsin E, TSLP, TCN2, GDF-15, epidermal morphogenetics, GRKS, PD-1, serine protease inhibitor A4, ADAM23, NOV, galactoglobulin-2, neuronal surface protein 3β, TLR3, Sirtuin2, Numb, IL-28 Rα, IL-33, Lin28, FCRL1, KLF4, NKp30, lymphocyte chemokine, cysteine ​​protease protein SN, JAM-A, calreticulin-2, ErbB4, BMP-8, IL-27 Ra, Fas, IL-4 Ra, kallikrein 14, extracellular matrix protein-3, Oligo2, kallikrein 12, CA13, IL-9, stalk protein-3, MPIF-1, cysteine ​​protease protein S, ADA, IL-2 Rb, GFR α-1, Smad4, ICAM-1, MEF2C, TREM-1, L-selectin, transmembrane serine protease, CD42b, MCSF, RANK, CHST4, CA8, FCRL3, ASAH2, CF XIV, PYY, HGF, I-TAC, brain signaling protein 4C, SorCS3, Tie-1, IL-31 RA, Arginase 1, POGLUT1, IL-1ra, Flatfoot protein, TIM-3, CREG, CD300f, uPA, EphA2, LLRTM4, LIMPII, Tenosynovin R, CPE, PECAM-1, DNAM-1, DKK-1, OPG, CPB1, TSH, MMP-2, Salivary lectin-9, ICAM-3, Cysteine ​​protease inhibitor SA, Galactohemagglutinin-4, Pepsinogen II, Desmosome core protein-3, Stalk protein-4, SCF, Serine protease inhibitor A5, PTH, FGF-19, MSP, IL-28A, FGF-12, METAP2, ASAHL, EDIL3, NTAL, EGF R, TAFAS, Galactohemagglutinin-9, vWF-A2, TACE, Activator protein RIM, Cathepsin S, LDL RBMPR-IA, OX40, IL-13 R2, B7-H4, MMP-13, ANGPTL7, TRAIL R4, IGSF4B, Sirtuin 5, PEAR1, SH2D1A, Cerberus 1, GDF-11, Nrf2, TROP-2, NUDTS, ROR2, EphB4, phosphatidylinositol proteoglycan 1, LAP (TGFb1), Gash, contactin-1, IL-27, UNC5H4, ICAM-2, MBL, HS3ST3B1, RCOR1, IL-10 Rb, XEDAR, IL-22, PILR-α, NRG1-131, FABP4, RGM-A, RELT, TrkC, CSa, SREC-I, neural epithelial stem cell protein, TPO, ErbB3, Kirrel3, FLRT1, galactoglobulin-3, CXCL16, JAM-B, DR6, Nogo receptor, TLR4, VEGF R2, Tie-2, IL-15 R, Caspr2, LTbR, LAMP, ALCAM, GLP-1, NG2, IL-22 Rα 1, AMIGO2, HCC-1, TFPI-2, ULBP-2, desmosome core protein 2, agglutinin, synaptic fusion protein 4, VAMP-1, stalkin-2, FGF-21, Flt-3, GFAP, TIM-1, inhibin A, cadherin-4, PIGF-2, neurogranulin, HE4, IL-23 R, galactoglobulin-7, GALNT3, GITR L, CD14, R-vertebral protein 2, CK19, cardiotrophin-1, TREML1, HAPLN1, CD27, ANG-4, salivary immunoglobulin-7, CD155, VEGF-C, TNF RII, PGRP-S, SDF-1a, PDGF-AB, GPVI, CD40, SCF R, platelet-reactive protein-5, IL-1 RII, neurofeedin-2, cadherin-13, E-selectin, GITR, WISP-1, renin, AgRP, MDL-1, ROBO3, RANTES, endothelial cell-specific molecules, granzyme, hCGb, mesothelin, TLR1, TRAIL, MOG, DDR1, NGF R, TRAIL R3, trypsin 3, ARSB, LIF Rα, BAFFR, CD157, granzyme A, 2B4, ESAM, IL-1 R4, CXCL14, IL-31, SIRP α, urinary modulatory proteins, CTRC, CEACAM-1, TARC, MIP-3a, SDF-1b, NKp46, MCP-3, IL-32 α, TGFb3, FOLR2, CD58, IL-23, CD36, TNFb, Shh-N, fibrinogen-1, Reg4ILT2, Mer, TREM-2, Flt-3L, CDS, IL-6, CD229, Insulin, Synaptic Fusion Protein 6, GRO, Bcl-w, Lipocrine-2, PDGF-AA, IL-2 Ra, Angiopoietin, LYVE-1, CD4, RAGE, CDNF, Short Proteoglycan, NAP-2, PU.1, EDAR, ADAMTS13, Kynureninase, PTH1R, IFN-γ R1, CrkL, B7-1, PARC, Draxin, VE-cadherin, Procalcitonin, SOX15, Kallikrein 11, BCMA, Dendritic Cell-Associated C-Lectin-2, EpCAM, HCC-4, TGFa, IP-10, BLAME, CILP-1, PIGF, LOX-1, MCP-2, Resistin, HVEM, ENPP-7, Cohesin-Glycan-4, IL-2 Rg, MICA, dopa decarboxylase, NPDC-1, MCP-4, EG-VEGF, glycoprotein V, brain signaling protein 4G, IL-12p40, total PSA, IL-15, MAP1D, C1q, TNF4, Dtk, endothelial glycoprotein, ENA-78, Reg3A, MIP-1b, FGF-17, IL-6R, IL-8, galactolectin-8, CA4, cysteine ​​protease protein EM, FUT8, B7-H 3. GCP-2, CD40L, MDC, 4-1BB, HO-1, SOST, S100A13, kallikrein 7 or IL-13 or a combination of two or more thereof; (ii) hsa-let-7a-5p, hsa-let-7b-5p, hsa-let-7c-5p, hsa-let-7d-3p, hsa-let-7e-5p, hsa-let-7g-5p, hsa-let-7i, hs a-let-7i-5p, hsa-miR-100-5p, hsa-miR-103a-3p, hsa-miR-106a-5p, hsa-miR-106b-5p, hsa-mir- 10b, hsa-miR-10b-5p, hsa-mir-1246, hsa-miR-1246, hsa-miR-125a-5p, hsa-miR-125b-5p, hsa-mi R-130a-3p, hsa-mir-130b, hsa-miR-130b-3p, hsa-miR-132-3p, hsa-miR-136-5p, hsa-miR-138-5p , hsa-miR-139-5p, hsa-mir-140, hsa-miR-140-3p, hsa-miR-145-5p, hsa-mir-146a, hsa-miR-146a- 5p, hsa-miR-148a-3p,hsa-miR-152-3p, hsa-miR-15a-5p, hsa-miR-15b-5p, hsa-mir-16-1, hsa-mir-16-2, hsa-miR-16-5p, hsa-miR-1'7-5p, hsa-miR-181a-5p, hsa-miR-191-5p, hsa-miR-193a-5p, hsa-miR-193b-3p, hsa-miR-19'7-3p, hsa-miR-199a-3p, hsa-miR-199a-5p, hsa-miR-199b-5p, hsa-miR-19a-3p, hsa-mi R-19b-3p, hsa-miR-20a-5p, hsa-mir-203a, hsa-miR-203a-3p, hsa-miR-214-3p, hsa-mir-21, hsa-miR-21-3p, hsa-miR-21-5p, hsa-mir-221, hsa-miR-221-3p, hsa-mir-222, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-mir-24-1, hsa-mir-24-2, hsa-miR-24-3p, hsa- mir-25, hsa-miR-25-3p, hsa-miR-26a-5p, hsa-miR-27a-3p, hsa-mir-27b, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-30a-5p, hsa-miR-30a-5p, hsa-miR-30b-5p, hsa-miR-30c-5p, hsa-mir-30d, hsa-miR-30d-5p, hsa-mir-30e, hsa-miR-30e-5p, hsa-miR-31-3p, hsa-miR-31-5p hsa-miR-320a hsa-miR-342-3p hsa-miR-345-5p hsa-miR-34a-5p hsa-miR-361-5p hsa-miR-376a-3p hsa-miR-376c-3p hsa-miR-423-3p hsa -miR-423-5p, hsa-miR-424-5p, hsa-miR-484, hsa-mir-486-1, hsa-mir-486-2, hsa-miR-486-5p, hsa-miR-570-3p, hsa-miR-574-3p, hsa-miR-663ahsa-miR-874-3p, hsa-mir-92a-1, hsa-mir-92a-2, hsa-miR-92a-3p, hsa-miR-92b-3p, hsa-mir-93, hsa-miR-93-5p, hsa-miR-940, hsa-miR-99a-5p or hsa-miR-99b-5p, or a combination of two or more thereof; or both (iii), (i) and (ii).

[0131] 45. Use of a composition comprising a therapeutic mesenchymal stem cell (MSC) secretome composition containing extracellular vesicles (EVs) in graft-versus-host disease in a therapeutic subject, wherein the composition comprises 15 mL of the therapeutic MSC secretome composition.

[0132] Example Example 1: Production of the therapeutic composition The MSC secretome therapeutic composition was prepared by the following method: Human bone marrow-derived MSCs were cultured in culture dishes with growth medium to expand the MSC population. The growth medium was then removed, and the cells were washed with PBS. The MSCs were then cultured under hypoxic conditions in a glucose-reduced medium with a pH less than 7.0. The conditioned medium was then collected and percolated, followed by sterilization. The production method of the therapeutic product was performed under current Good Manufacturing Practice and Current Good Tissue Practice.

[0133] The tetraspan membrane protein profile of extracellular vesicles present in the therapeutic composition was determined, and it was found that more than 95% of the extracellular vesicles present in the therapeutic composition were CD63. + CD9 - CD81 - .

[0134] The protein contents of the therapeutic product were identified, and the following proteins were found: ferritin, IGFBP-4 (insulin-like growth factor binding protein-4), IL-1 R6 (interleukin-1 receptor 6), LAMP2 (lysosome-associated membrane glycoprotein 2), bIG-H3 (transforming growth factor-β-inducible protein ig-h3), GPR115 (adhesion G protein-coupled receptor F4), CD63 antigen, CD109 antigen, serine protease inhibitor F1 (pigment epithelium-derived factor), IGFBP-6 (insulin-like growth factor binding protein-6), HS3ST4 (heparanol sulfate glucosamine 3-O-sulfotransferase 4), OPN (osteopontin), and PAI-1 (plasminogen activator inhibitor-1 or sERPINE). 1) Cathepsin B, IGFBP-2 (insulin-like growth factor binding protein-2), brain signaling protein 6C, IGF-2 (insulin-like growth factor-2), sorting protein, serine protease inhibitor B6, Dkk-3 (Dickkopf-related protein 3), CNTF (ciliary neurotrophic factor), TSP-1 (thromboretin-1), GM-CSF Ra (granulocyte-macrophage colony-stimulating factor receptor subunit α), coagulation regulatory protein, endoglucan, (podocyte protein-like protein 2) IGFBP-3 (insulin-like binding protein-3), RGM-C (hepcidin regulatory protein), PF4 (platelet factor 4), MIF (macrophage migration inhibitory factor), TGM4 (protein-glutamine-γ-glutamyltransferase 4), periosteal protein, furin protease, TIMP-1 (MMP tissue inhibitor 1), gypsum glycan, PCK1 (cytosol-type phosphoenolpyruvate carboxykinase), CD9 antigen, CD99 antigen, CA2 (carbonic anhydrase 2), PRDX4 (peroxide reductase-4), transferrin, DcR3 (tumor necrosis factor receptor superfamily member 6B), GP73 (Golgi membrane protein 1), CD81 antigen, phytoglucan, and TIMP-2 (MMP tissue inhibitor 2).

[0135] The nucleic acid inclusions of the therapeutic product were identified, and the following nucleic acids were found to be present: hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-21-5p, hsa-miR-221-3p, hsa-miR-222-3p, hsa-miR-22-3p, and hsa-miR-23a-3p. , hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa- miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p and hsa-miR-940.

[0136] Example 2: Treatment in humans Regulatory approval Following FDA approval of an Investigational New Drug (IND) application for a single patient, informed consent is obtained from the patient, and treatment is administered. Institutional Review Board (IRB) approval is obtained. Each patient has a unique treatment plan submitted to the FDA, but all include the initial three doses of the investigational treatment product, 15 mL every other day for seven days.

[0137] Therapeutic products The investigational therapeutic product administered was an EV product derived from bone marrow MSCs from a single donor and manufactured according to stringent quality standards (with US FDA master file) and in accordance with US FDA GMP regulations to ensure batch quality and consistency. The MSC-derived EV product was produced from bone marrow MSCs cultured under hypoxic, hypoglycemic, and hypopH conditions. For example, the product from Example 1.

[0138] Dosage and safety Previous studies have demonstrated the safety of IV doses with capped doses of 1 million cells / kg, 5 million cells / kg, and in addition, 10 million cells / kg; (3) approximately 2,000 extracellular vesicles were observed secreted per stromal cell; and (4) laboratory analysis of the IP showed approximately 60-80 billion extracellular vesicles per mL. For a 70 kg adult, extrapolating the MSC capped dose from the START trial yields a capped dose of 17.5 mL of IV therapeutic product. Taking into account the range of body mass index (BMI), 15 mL of IV therapeutic product was determined as the starting dose for each infusion. Based on FDA-approved clinical trials, no serious adverse events were attributed to the use of the therapeutic product within this dose range. Based on previous preclinical studies of extracellular vesicles derived from human bone marrow, there was also no known possibility of malignant transformation.

[0139] Patient presentation The demographic characteristics and transplant history of the patients are shown in Table 2 below. Three patients had modified multi-organ allogeneic grafts, two had separate intestinal allogeneic grafts, and two had separate liver allogeneic grafts; six were adults and one was a pediatric patient.

[0140] Table 2 Treatment goals The goal of this emergency use treatment is to administer this therapeutic product in life-threatening situations due to GVHD complicated by pulmonary involvement and / or signs of allogeneic graft rejection despite high-dose corticosteroids. The primary goal is to assess safety by monitoring for any adverse events and evidence of injection toxicity. Secondary goals are clinical, serological, and histopathological evidence of improvement in allogeneic graft rejection and signs of GVHD.

[0141] Treatment plan and administration method All patients continued their immunosuppressive medications during the treatment regimen (unless they had lost a response or had contraindications) (see Table 2). The treatment plan submitted to the FDA, specifying the total number of doses for each patient and the dose volume for pediatric patients, was based on the adult and pediatric patient's condition, the severity of the current disease, the stage of acute or chronic disease, and the presence of an allogeneic graft. The general treatment plan included: an intravenous infusion of 15 mL of the treatment product suspended in a total of 100 mL of normal saline on day 0. The 100 mL was infused over 1 hour without a filter. This procedure was repeated on days 2 and 4 after assessment confirming signs of infusion toxicity without adverse events. Patients' clinical status, serology, and histopathology were assessed post-infusion according to standard of care (Table 3).

[0142] Table 3 result Acute monitoring after therapeutic product delivery Patients were observed in the post-infusion unit after administration of the therapeutic product to assess acute adverse events, response to the product, or significant changes in vital signs.

[0143] Clinical assessment No adverse or serious adverse events related to the treatment product occurred during the treatment program in all seven patients. All patients experienced improvement in clinical symptoms within 24 hours of the initial 15 mL treatment product dose. Similarly, in those patients with graft-versus-host disease (n=2), pulmonary symptoms and dermatological manifestations of graft-versus-host disease improved within 24 hours of the initial 15 mL treatment product dose. All patients had improved serological laboratory assessments related to transplant function within 7 days of their initial treatment product dose and achieved complete histological resolution of graft inflammation / rejection within 7 days of treatment (Tables 2 and 3).

[0144] Figure 1The following images show histological improvement in inflammation and resolution of acute cellular rejection following treatment product administration. (A) Severe acute cellular rejection of the intestinal allogeneic graft in Patient 1, associated with confluent crypt cell apoptosis, crypt loss, and ulceration. (B) Resolution of acute cellular rejection of the intestinal allogeneic graft in Patient 1 after treatment product administration. (C) Severe acute cellular rejection of the liver allogeneic graft in Patient 2, characterized by extensive bile duct injury, portal endotheliitis, and central hepatocellular injury. (D) Significant histological improvement of the liver allogeneic graft in Patient 2 after treatment product administration, with only focal residual cholangitis. (E) Mild acute cellular rejection of the intestinal allogeneic graft in Patient 3, characterized by increased apoptotic crypt epithelial cells. (F) Complete histological resolution of acute cellular rejection in Patient 3. (G) Severe acute cellular rejection of the intestinal allogeneic graft in Patient 4, with crypt loss and ulceration. (H) Patient 4 showed complete mucosal healing of the intestinal allogeneic graft after treatment product administration, with no residual apoptotic activity or increased crypt loss. (I) Patient 5 experienced mild acute cellular rejection of the intestinal allogeneic graft, with only occasional increase in apoptotic crypt epithelial cells. (J) Patient 5 experienced resolution of acute cellular rejection after treatment product administration. (K, L) Patient 6 showed normal small intestinal allogeneic graft before and after treatment with the treatment product for GVHD.

[0145] In summary, the safety and efficacy of extracellular vesicle (EV) products derived from mesenchymal stem cells were tested in a transplant patient population. Seven independent Emergency Investigational New Drug (eIND) applications were submitted to the FDA for the emergency treatment of graft-versus-host disease (GVHD) in isolated intestinal grafts (n=2), liver allogeneic grafts (n=2), modified multi-organ grafts (n=3), and isolated intestinal transplant patients (n=2). Following FDA-approved treatment protocols, 15 mL of the treatment product was administered intravenously on days 0, 2, and 4, and this treatment cycle was repeated up to four times per patient. Safety (adverse event reporting) and efficacy (clinical status, serology, and histopathology) were evaluated. No adverse events were associated with the treatment product. All patients experienced improvement in clinical symptoms, serological laboratory assessments, pulmonary symptoms and dermatological manifestations of GVHD within 24 hours of treatment product administration, and complete histological resolution of graft inflammation / rejection within 7 days of treatment product administration. The systemic application of MSC-derived EV products has successfully achieved histological clearance of graft inflammation in the intestine, liver, and multiple organs, as well as skin and lung manifestations of GVHD.

[0146] Example 3: Characterization of Improved Serum Inflammatory Markers Patients are treated with the therapeutic product of this disclosure as described in Example 2. In some embodiments, patients treated with the therapeutic product have severe or life-threatening abdominal solid organ transplant rejection, or are assessed and determined to be at high risk of progressing to severe or life-threatening conditions associated with abdominal solid organ transplant rejection, at risk of allogeneic graft dysfunction, or at risk of complications from current immunosuppressive therapy regimens. Patients are assessed before, during, and after treatment to characterize improvements seen in serum inflammatory markers and inflammation of solid abdominal organs. This may include improvements seen in the following transplant types: • Liver transplantation: Overall improvement in liver function, as seen through reduced liver function tests and improved albumin levels.

[0147] • Small bowel transplantation: Biopsy showed reduced inflammation • Pancreas: Decreased amylase and lipase levels • Multiple organs: any of the above criteria Over a period of one year, patients were given 15 mL of the treatment product intravenously in a total volume of 100 mL (mixed with 85 mL of normal saline), up to nine times. Each patient was followed up for 6 months after treatment. The treatment product of Example 1 is used as an example.

[0148] Inclusion criteria: • Provide a signed and dated informed consent form. • Demonstrates willingness to comply with all research procedures and participate throughout the research period. • Male or female, aged 18 to 75 • Previous abdominal solid organ transplant • Diagnosis of acute or chronic rejection based on clinical observation, laboratory analysis, and histological evaluation.

[0149] • Diagnosis of deteriorated allogeneic graft function based on clinical observation, laboratory analysis, and histological assessment.

[0150] • Failure of first-line and alternative standard care therapies • According to the attending physician's assessment, the patient has a serious or life-threatening condition, or a condition that will progress to a serious or life-threatening condition if left untreated.

[0151] • If medical history indicates that overall health is good. • Men or women of fertility potential must use fertility control measures from enrollment until 6 months after the study is completed. Exclusion criteria: • Pregnancy or breastfeeding • Receive another investigational drug or other intervention within 30 days of enrollment. • Subjects with a history of laboratory evidence of hypercoagulable states • Individuals with a history of deep vein thrombosis or pulmonary embolism.

[0152] While preferred embodiments of this disclosure have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Various modifications, alterations, and substitutions will now be made by those skilled in the art without departing from this disclosure. It should be understood that various alternatives to embodiments of this disclosure may be employed in the practice of this disclosure. The following claims are intended to define the scope of this disclosure and thereby cover the methods and structures within the scope of these claims and their equivalents.

Claims

1. A method for treating graft-versus-host disease in a subject in need, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs); wherein the one or more EVs comprise hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-21-5p, hsa-miR-221-3p, hsa-miR-222-3p, hsa-miR-22- 3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, or hsa-miR-940, or a combination of two or more thereof.

2. A method for treating graft-versus-host disease in a subject in need, the method comprising administering to the subject a composition comprising one or more extracellular vesicles (EVs); wherein at least 80% of the EVs are CD63+, CD9-, or CD81-.

3. A method for treating graft-versus-host disease in a recipient, the method comprising administering to the recipient a composition comprising: ferritin, IGFBP-4 (insulin-like growth factor binding protein-4), IL-1 R6 (interleukin-1 receptor 6), LAMP2 (lysosome-associated membrane glycoprotein 2), bIG-H3 (transforming growth factor-β-inducible protein ig-h3), GPR115 (adhesion G protein-coupled receptor F4), CD63 antigen, CD109 antigen, serine protease inhibitor F1 (pigment epithelial-derived factor), IGFBP-6 (insulin-like growth factor binding protein-6), HS3ST4 (heparanol sulfate glucosamine 3-O-sulfotransferase 4), OPN (osteopontin), PAI-1 (plasminogen activator inhibitor-1 or sERPINE). 1) Cathepsin B, IGFBP-2 (insulin-like growth factor binding protein-2), brain signaling protein 6C, IGF-2 (insulin-like growth factor-2), sorting protein, serine protease inhibitor B6, Dkk-3 (Dickkopf-related protein 3), CNTF (ciliary neurotrophic factor), TSP-1 (thromboretin-1), GM-CSF Ra (granulocyte-macrophage colony-stimulating factor receptor subunit α), coagulation regulatory protein, endosaccharide, (podocyte protein-like protein 2) IGFBP-3 (insulin-like binding protein-3), RGM-C (hepcidin regulatory protein), PF4 (platelet factor 4), MIF (macrophage migration inhibitory factor), TGM4 (protein-glutamine-γ-glutamyltransferase 4), periosteal protein, furin protease, TIMP-1 (MMP tissue inhibitor 1), gypsum proteoglycan, PCK1 (cytosol-type phosphoenolpyruvate carboxykinase), CD9 antigen, CD99 antigen, CA2 (carbonic anhydrase 2), PRDX4 (peroxide reductase-4), transferrin, DcR3 (tumor necrosis factor receptor superfamily member 6B), GP73 (Golgi membrane protein 1), CD81 antigen, luminescent proteoglycan, or TIMP-2 (MMP tissue inhibitor 2), or a combination of two or more thereof.

4. The method of claim 2, wherein the one or more EVs comprise hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-21-5p, hsa-miR-221-3p, hsa-miR-222-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa- miR-23b-3p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, or hsa-miR-940, or a combination of two or more thereof.

5. The method of claim 1 or 2, wherein the composition comprises ferritin, IGFBP-4 (insulin-like growth factor binding protein-4), IL-1 R6 (interleukin-1 receptor 6), LAMP2 (lysosome-associated membrane glycoprotein 2), bIG-H3 (transforming growth factor-β inducible protein ig-h3), GPR115 (adhesion G protein-coupled receptor F4), CD63 antigen, CD109 antigen, serine protease inhibitor F1 (pigment epithelial-derived factor), IGFBP-6 (insulin-like growth factor binding protein-6), HS3ST4 (heparanol sulfate glucosamine 3-O-sulfotransferase 4), OPN (osteopontin), PAI-1 (plasminogen activator inhibitor-1 or sERPINE). 1) Cathepsin B, IGFBP-2 (insulin-like growth factor binding protein-2), brain signaling protein 6C, IGF-2 (insulin-like growth factor-2), sorting protein, serine protease inhibitor B6, Dkk-3 (Dickkopf-related protein 3), CNTF (ciliary neurotrophic factor), TSP-1 (thromboretin-1), GM-CSF Ra (granulocyte-macrophage colony-stimulating factor receptor subunit α), coagulation regulatory protein, endosaccharide, (podocyte protein-like protein 2) IGFBP-3 (insulin-like binding protein-3), RGM-C (hepcidin regulatory protein), PF4 (platelet factor 4), MIF (macrophage migration inhibitory factor), TGM4 (protein-glutamine-γ-glutamyltransferase 4), periosteal protein, furin protease, TIMP-1 (MMP tissue inhibitor 1), gypsum proteoglycan, PCK1 (cytosol-type phosphoenolpyruvate carboxykinase), CD9 antigen, CD99 antigen, CA2 (carbonic anhydrase 2), PRDX4 (peroxide reductase-4), transferrin, DcR3 (tumor necrosis factor receptor superfamily member 6B), GP73 (Golgi membrane protein 1), CD81 antigen, luminescent proteoglycan, or TIMP-2 (MMP tissue inhibitor 2), or a combination of two or more thereof.

6. The method of any one of claims 1-5, wherein the subject has received an organ transplant or allogeneic graft.

7. The method of claim 6, wherein the subject has received a modified multi-organ allogeneic graft, a single intestinal allogeneic graft, or a single liver allogeneic graft.

8. The method of any one of claims 1-7, wherein the subject exhibits one or more signs or symptoms of graft-versus-host disease and / or allogeneic graft rejection.

9. The method of any one of claims 1-8, wherein the subject exhibits one or more signs or symptoms selected from the following: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, itching, spasms, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof.

10. The method of any one of claims 1-9, wherein the subject is receiving corticosteroids.

11. The method of claim 10, wherein the subject is receiving a high dose of corticosteroids.

12. The method of any one of claims 1-11, wherein the subject experiences improvement in one or more signs or symptoms selected from the following after treatment: crypt cell apoptosis, crypt loss, ulceration, cell rejection of allogeneic grafts, bile duct injury, portal vein endotheliitis, central hepatocellular injury, cholangitis, inflammation, pain, bleeding, fever, chills, redness, burning sensation, pruritus, spasms, nausea, vomiting, loss of appetite, jaundice, hepatomegaly, rash, vesicles, peeling, tenderness, liver failure, ulceration, and combinations thereof.

13. The method of any one of claims 1-12, wherein the dose of the therapeutic MSC secretory composition administered to the subject is a cell equivalent dose of 700,000 to 7,000,000 cells / kg.

14. The method of any one of claims 1-13, wherein administration includes intravenous administration.

15. The method of any one of claims 1-14, wherein the composition is prepared by a method comprising the steps of: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under conditions of: (i) oxygen tension below 5%; and (ii) a medium with a pH below 7; (b) harvesting the MSC conditioned medium; and (c) formulating the MSC conditioned medium to produce the composition.

16. The method of any one of claims 1-15, the method comprising preparing the composition prior to the administration, wherein the preparation of the composition is carried out by a method comprising the steps of: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under conditions of: (i) oxygen tension below 5%; and (ii) a medium with a pH below 7; (b) harvesting the MSC conditioned medium; and (c) preparing the MSC conditioned medium to produce the composition.

17. The method of claim 15 or 16, wherein the culture medium is serum-free.

18. The method of any one of claims 15-17, wherein the culture medium has a glucose concentration of less than 4.5 g / L.

19. The method of any one of claims 15-18, wherein preparing the MSC conditioned medium comprises replacing the conditioned medium with a pharmaceutically acceptable formulation.

20. The method of claim 19, wherein the pharmaceutically acceptable formulation comprises saline solution.

21. The method of any one of claims 1, 2, or 4, wherein the composition comprises at least 6 × 10 10 Up to 8 × 10 10 One extracellular vesicle per ml, and administered at doses of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 ml.

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