Mesenchymal stem cell compositions and methods of making

Abstract

Disclosures herein are directed to first compositions comprising exosomes and extracellular matrix components and their use thereof. Also described are methods and kits wherein these first compositions are packaged and used with second compositions comprising mesenchymal stem cells. The first and second compositions are derived from the same tissue source using tangential flow filtration.

What is claimed is: 
     
         1 . An exosome composition comprising (a) at least one exosome produced by a mesenchymal stem cell or fibroblast cell, (b) at least one component isolated from an extracellular matrix (an ECM component). 
     
     
         2 . The exosome composition of  claim 1 , wherein the at least one ECM component is selected from the group consisting of a glycosaminoglycan or proteoglycan thereof, an adhesive glycoprotein, a fibrous protein, a growth factor, a cytokine, a peptide, a nucleic acid, or a lipid. 
     
     
         3 . The exosome composition of  claim 1 , wherein the at least one ECM component comprises; (a) a glycosaminoglycan selected from the group consisting of hyaluronan, chondroitin sulfate (CS), dermatan sulfate (DS), keratan sulfate (KS), and heparan sulfate (HS), or any proteoglycan thereof, (b) an adhesive glycoprotein selected from the group consisting of Laminin, Fibronectin, tenascin, and nidogen, (c) a fibrous protein selected from the group consisting of collagens and elastin, and/or (d) a growth factor selected from the group consisting of EGF, Endothelin, Eotaxin/CCL11, FGF-4, GDF-15, ICAM-1, IGFBP-1, IL-6, PDGF-AA, TGF-beta 3, TIMP-1, TIMP-2, BCAM, Smad 4, CD 163, CD30 Ligand, TNFSF8, and Integrin b1. 
     
     
         4 . (canceled) 
     
     
         5 . (canceled) 
     
     
         6 . (canceled) 
     
     
         7 . (canceled) 
     
     
         8 . The exosome composition of  claim 1 , wherein the at least one ECM component is present at a higher concentration than a concentration of the ECM component in a comparable exosome preparation. 
     
     
         9 . (canceled) 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . (canceled) 
     
     
         14 . The exosome composition of  claim 8 , wherein the concentration of the ECM component is at least 20 times the concentration of the ECM component in a comparable exosome preparation. 
     
     
         15 . The exosome composition of  claim 14 , wherein the ECM component is selected from the group consisting of: Cysteine-rich Protein 1, PIM2, Cadherin-13, Beta IG-H3, EG-VEGF/PK1, Cyclophilin A, Dkk-3, BMP-3, QDPR, PAM, EDAR, EGF R/ErbB1, ALCAM, CXCR3, SBP-1, BCMA/TNFRSF17, CCL28/VIC, IL-29, HSP47, VE-Cadherin, I-309, TGF-beta RIII, CCR8, Fractalkine, TCN1, RSU1, uPAR, VEGF-D, PRELP, TGF-beta RII, Thrombopoietin (TPO), Endothelin, S100A6, Angiogenin, TIMP-4, MSP alpha Chain, Activin A, DMGDH, Serpin B8, TRAIL R3/TNFRSF10C, CFHR5, Caspase-14, PRDX 1, FGF-12, CXCR5/BLR-1, Semaphorin 7A, SPINK7, NT-4, TNF RII/TNFRSF1B, 6Ckine, SerRS, Angiostatin, Angiopoietin-like Factor, B7-1/CD80, Protein C, Tcf20, TROY/TNFRSF19, EMAP-II, CSH1, TLR4, RPLP0, ErbB3, RPL11, PGK-1, FGF-16, Plakophilin 1, PRCP, RAGE, Insulin, TSR2, NT-3, USP14, VDAC1/Porin, Proteasome 20S a+b, M-CSF, IL-2, ICAM-1, NEDD8, PRG2, Insulysin/IDE, ICAM-2, MMP-1, Salivary alpha amylase/aAmylase, IL-17F, Galectin-1, Lymphotoxin beta/TNFSF3, TBCA, PREP, VAP-A, Utrophin, YB1, VAP-1, CFHR4, FGF-20, Cyclophilin B, RNASE4, SBSN, FGF R4, FGF-17, PGLS-C-t, Eotaxin-3/CCL26, RREB1, TMEM223, ADAMTS-15, Semenogelin II/SEMG2, C3a, RPL12, GDF8, IGF-II, RPS25, Follistatin-like 1, Smad 5, Notch-2, I-TAC/CXCL11, TRANCE, MMP-3, uPA, WDR1, UQCRB, DR6/TNFRSF21, BMP-15, PSMA4, ErbB4, Gephyrin, beta-NGF, BMP-2, BMP-6, NAP-2, LECT2, FGF-13 1B, Reg3A, CD14, SHIP, RPL5, VEGF, IL-1 F5/FIL1delta, RPS28, TUBA6, Endostatin, MMP-9, CK-MB, CapG, IL-11, Ribonuclease A, Eotaxin-2/MPIF-2, GLO-1, TLR3, HGF, Sterol carrier protein 2/SCP2, Visfatin, SHC1, IL-18 R beta/AcPL, SIGLEC14, PRTN3, Contactin-1, CV-2/Crossveinless-2, EDG-1, CXCL14/BRAK, TIMP-3, Serpin A1, Annexin A1, TGM3, CXCR6, PGM1, Osteoprotegerin/TNFRSF11B, Coagulation Factor III/Tissue Factor, M-CSF R, MDC, FAM3B, MMP-11/Stromelysin-3, SCF R/CD 117, TGF-beta 5, Titin, sFRP-3, IL-1 beta, HCC-4/CCL16, TRAIL/TNFSF10, Fascin, BNP, ADAMTS-13, PLOD1, FLRG, LDHA, LIF, YY1, NME3, ESAM, RPS19, EpCAM, TAGLN2, CXCL16, BTF3, Hemoglobin subunit beta/HBB, TRADD, Metavinculin, BDNF, Galectin-7, PPP2R4, FACX, PDZD2, IL-26, FGF-9, Fibrinogen-like 2, Plexin B2, Thrombomodulin, Dtk, TRAIL R2/DR5/TNFRSF10B, PNP, ANGPTL8, B7-H3, ACLP, Galectin-3, p73, IL-4, IL-13 R alpha 2, TOB2, Claudin-3, IL-8, VEGF-C, IGF-I SR, Tenascin X(1), Lefty-A, Thrombospondin-1, OSM R beta, SUCLG1, PDGF-AB, IL-6 R, IL-18 BPa, PGAM2, Cripto-1, FGF-23, GSTM1, ICAM-5, RPL10, Ribonuclease Inhibitor, ARTS1, Periostin, UCH-L1, Zyxin, RPL10A, PDLIM5, IL-15 R alpha, FAM3C, Tropomyosin 3, LZTS1, LIGHT/TNFSF14, Osteoadherin(2), SHMT1, CCR9, TWEAK R/TNFRSF12, TPP1, ADAMTS-10, RCL, PCDH7, MIG, SPARC, SH3BGRL, Thrombospondin-4, UFM 1, Prostaglandin D Synthase/PTGDS, GPR-39, SPTBN1, BLVRB, RKIP, 4-1BB, Decorin, CRIM 1, PCSK9, STI1, IL-22 BP, VEGF R2 (KDR), Ficolin-2, CXCR4 (fusin), SIGIRR, IFN-beta, Cyclin D1, MIP-1a, RPS11, Alpha 1 AG, BLAME, Cystatin E/M, GM-CSF R alpha, Uteroglobin(1), Trypsinogeb-2, MCP-3, Erythropoietin R, Proteasome subunit alpha type 6/PSMA6, TGF-alpha, Flt-3 Ligand, Nanog, TNF-alpha, Spectrin beta-5, VNN1, BMP-1, FGF-7/KGF, OSM, FGF R5, and Transferrin. 
     
     
         16 . The exosome composition of  claim 8 , wherein the concentration of the ECM component is at least 30 times the concentration of the ECM component in a comparable exosome preparation. 
     
     
         17 . The exosome composition of  claim 15 , wherein the ECM component is selected from the group consisting of Cysteine-rich Protein 1, PIM2, Cadherin-13, Beta IG-H3, EG-VEGF/PK1, Cyclophilin A, Dkk-3, BMP-3, QDPR, PAM, EDAR, EGF R/ErbB1, ALCAM, CXCR3, SBP-1, BCMA/TNFRSF17, CCL28/VIC, IL-29, HSP47, VE-Cadherin, I-309, TGF-beta RIII, CCR8, Fractalkine, TCN1, RSU1, uPAR, VEGF-D, PRELP, TGF-beta RII, Thrombopoietin (TPO), Endothelin, S100A6, Angiogenin, TIMP-4, MSP alpha Chain, Activin A, DMGDH, Serpin B8, TRAIL R3/TNFRSF10C, CFHR5, Caspase-14, PRDX 1, FGF-12, CXCR5/BLR-1, Semaphorin 7A, SPINK7, NT-4, TNF RII/TNFRSF1B, 6Ckine, SerRS, Angiostatin, Angiopoietin-like Factor, B7-1/CD80, Protein C, Tcf20, TROY/TNFRSF19, EMAP-II, CSH1, TLR4, RPLP0, and ErbB3. 
     
     
         18 . The exosome composition of  claim 8 , wherein the concentration of the ECM component is at least 40 times the concentration of the ECM component in a comparable exosome preparation comprising an intact extracellular matrix. 
     
     
         19 . The exosome composition of  claim 18 , wherein the ECM component is selected from the group consisting of Cysteine-rich Protein 1, PIM2, Cadherin-13, Beta IG-H3, EG-VEGF/PK1, Cyclophilin A, Dkk-3, BMP-3, QDPR, PAM, EDAR, EGF R/ErbB1, ALCAM, CXCR3, and SBP-1. 
     
     
         20 . (canceled) 
     
     
         21 . (canceled) 
     
     
         22 . The exosome composition of  claim 8 , wherein the comparable exosome preparation is an exosome preparation prepared via ultra-centrifugation. 
     
     
         23 . The exosome composition of  claim 1 , comprising 1 billion or more exosomes. 
     
     
         24 . The exosome composition of  claim 1 , further comprising a carrier or excipient. 
     
     
         25 . (canceled) 
     
     
         26 . (canceled) 
     
     
         27 . A method for preparing an exosome composition and an MSC composition, the method comprising: (a) culturing mesenchymal stem cells isolated from a tissue sample in an exosome collection media, (b) collecting the exosome collection media as a first fraction and the mesenchymal stem cells as a second fraction, the second fraction further comprising exosomes and extracellular matrix colonies and fragments thereof, (c) filtering the second fraction through a first section of a tangential flow filtration system to obtain a retentate comprising mesenchymal stem cells and a first filtrate comprising exosomes and the extracellular matrix colonies and fragments thereof, (d) collecting the retentate comprising mesenchymal stem cells as a MSC composition, (e) filtering the first filtrate through a second section of the tangential flow filtration system to form a second retentate comprising the and the extracellular matrix colonies and fragments thereof and a second filtrate comprising exosomes and at least one component from the extracellular matrix, (f) combining the second filtrate with the first fraction to form a third fraction, and (g) collecting the third fraction as the exosome composition. 
     
     
         28 . (canceled) 
     
     
         29 . (canceled) 
     
     
         30 . (canceled) 
     
     
         31 . (canceled) 
     
     
         32 . (canceled) 
     
     
         33 . (canceled) 
     
     
         34 . (canceled) 
     
     
         35 . (canceled) 
     
     
         36 . (canceled) 
     
     
         37 . (canceled) 
     
     
         38 . (canceled) 
     
     
         39 . A kit comprising:
 (a) the exosome composition of  claim 1 ; or;   (b) an MSC composition comprising one or more mesenchymal stem cells, wherein the MSC composition is free of extracellular matrix colonies and fragments thereof; or   (a) and (b).   
     
     
         40 . The kit of  claim 39 , wherein the kit comprises (a). 
     
     
         41 . The kit of  claim 39 , wherein the kit comprises (b). 
     
     
         42 . The kit of  claim 39 , wherein the kit comprises (a) and (b). 
     
     
         43 . A method of treating a subject in need thereof, comprising administering the exosome composition of  claim 1 , and at least one mesenchymal stem cell to the subject. 
     
     
         44 . The method of  claim 43 , wherein administering at least one mesenchymal stem cell comprises administering an MSC composition comprising one or more mesenchymal stem cells, wherein the MSC composition is free of extracellular matrix colonies and fragments thereof to the subject.