Reduction of senescent cell associated aging and pathology

Abstract

Compositions of matter, devices, and treatment protocols for reducing deleterious effects of senescent cells in a mammal. The invention provides extracorporeal devices capable of removing, in some cases selectively, senescence associated factors including small molecules, proteins, lipoproteins, as well as in some embodiments extracellular vesicles including exosomes, apoptotic bodies, or cellular released particles. Extracorporeal removal of senescent cell associated factors is performed to augment effects of senolytic therapy. Removal of senescence associated factors is performed to enhance efficacy of regenerative medicine interventions.

Description

REDUCTION OF SENESCENT CELL ASSOCIATED AGING AND PATHOLOGYCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to United States Provisional Application No. 63 / 673,915, filed on July 22, 2024, the contents of which are incorporated herein by reference.FIELD OF THE INVENTION

[0002] The invention pertains to the field of experimental gerontology, more specifically, the invention pertains to interventional gerontology, more specifically, the invention pertains to removal of senescent associated factors extracorporeally in order to induce regenerative effects and / or enhance therapeutic effects of regenerative interventions.BACKGROUND

[0003] Senescence occurs after proliferative exhaustive and / or accumulation of molecular damage. Senescent cells accumulate in tissues and organs of individuals as they age and are found at sites of age-related pathologies. While senescent cells are believed important to inhibiting proliferation of dysfunctional or damaged cells and particularly to constraining development of malignancy, the presence of senescent cells in an aging individual may contribute to aging and aging-related dysfunction. Given that senescent cells have been causally implicated in certain aspects of age-related decline in health and may contribute to certain diseases, and are also induced as a result of necessary life-preserving chemotherapeutic and radiation treatments, the presence of senescent cells may have deleterious effects to millions of patients worldwide (e.g., fatigue, weakness, loss of physical agility, decrease in cognitive function). Accordingly, treatments aimed at clearing aging-induced and therapy-induced senescent cells and improving age-sensitive traits have the potential to markedly improve the health, lifespan, and quality of life for patients exposed to senescence-inducing stimuli. The present disclosure addresses these needs and offers numerous related advantages.

[0004] Senescent cells are known to secrete multiple factors that are detrimental to the host and in some cases produce an “infectious senescence”. These factors, called“senescent associated secretory phenotype” (SASPS) include inflammatory cytokines such as IL-6, IL-1 beta and TNF-alpha. In numerous disease conditions, senescent cells have been demonstrated to play a deleterious role ranging from intervertebral disc degeneration, to liver failure, to cancer, to heart failure.SUMMARY

[0005] Compositions of matter, devices, and treatment protocols for reducing deleterious effects of senescent cells in a mammal. In one embodiment the invention provides extracorporeal devices capable of removing, in some cases selectively, senescence associated factors including small molecules, proteins, lipoproteins, as well as in some embodiments extracellular vesicles including exosomes, apoptotic bodies, or cellular released particles. In some embodiments extracorporeal removal of senescent cell associated factors is performed to augment effects of senolytic therapy. In other embodiments removal of senescence associated factors is performed to enhance efficacy of regenerative medicine interventions.

[0006] Various aspects of the invention are enumerated in the following paragraphs:

[0007] Aspect 1 : A method of reducing pathological effects of senescent cell accumulation in a subject, comprising: a) Identifying a patient with senescent cell accumulation; b) Quantifying senescence or senescence-associated biomarkers; c) Extracorporeally removing senescence-associated secretory factors; and d) Optionally, re-assessing said senescence-associated biomarkers to determine the need for additional extracorporeal removal.

[0008] Aspect 2: The method of Aspect 1, wherein said senescent cells are metabolically active.

[0009] Aspect 3: The method of Aspect 1, wherein said senescent cells exhibit enhanced p!6 expression compared to non-senescent cells.

[0010] Aspect 4: The method of Aspect 1, wherein said senescent cells are in cell cycle arrest.

[0011] Aspect 5: The method of Aspect 1, wherein said senescent cells exhibit enhanced p53 expression compared to non-senescent cells.

[0012] Aspect 6: The method of Aspect 1, wherein said senescent cells produce at least 20% more interleukin-1 beta compared to non-senescent cells.

[0013] Aspect 7: The method of Aspect 1, wherein said senescent cells produce at least 50% more interleukin-1 beta compared to non-senescent cells.

[0014] Aspect 8: The method of Aspect 1, wherein said senescent cells produce at least 100% more interleukin- 1 beta compared to non-senescent cells.

[0015] Aspect 9: The method of Aspect 1, wherein said senescent cells produce at least 20% more interleukin-6 compared to non-senescent cells.

[0016] Aspect 10: The method of Aspect 1, wherein said senescent cells produce at least 50% more interleukin-6 compared to non-senescent cells.

[0017] Aspect 11 : The method of Aspect 1, wherein said senescent cells produce at least 100% more interleukin-6 compared to non-senescent cells.

[0018] Aspect 12: The method of Aspect 1, wherein said senescent cells produce at least 20% more interleukin-8 compared to non-senescent cells.

[0019] Aspect 13: The method of Aspect 1, wherein said senescent cells produce at least 50% more interleukin-8 compared to non-senescent cells.

[0020] Aspect 14: The method of Aspect 1, wherein said senescent cells produce at least 100% more interleukin-8 compared to non-senescent cells.

[0021] Aspect 15: The method of Aspect 1, wherein said senescent cells exhibit the Senescence- Associated Secretory Phenotype (SASP).

[0022] Aspect 16: The method of Aspect 1, wherein said SASP is associated with enhanced inflammasome activation in cells surrounding said senescent cells.

[0023] Aspect 17: The method of Aspect 1, wherein said SASP is associated with enhanced NF-kappa B activation in cells surrounding said senescent cells.

[0024] Aspect 18: The method of Aspect 1, wherein said SASP is associated with enhanced Rel-B activation in cells surrounding said senescent cells.

[0025] Aspect 19: The method of Aspect 1, wherein said SASP is associated with enhanced Rel-C activation in cells surrounding said senescent cells.

[0026] Aspect 20: The method of Aspect 1, wherein said SASP is associated with enhanced p38 MAPK activation in cells surrounding said senescent cells.

[0027] Aspect 21 : The method of Aspect 1, wherein said SASP is associated with enhanced JAK-STAT activation in cells surrounding said senescent cells.

[0028] Aspect 22: The method of Aspect 1, wherein said SASP is associated with enhanced STING activation in cells surrounding said senescent cells.

[0029] Aspect 23: The method of Aspect 1, wherein said SASP is associated with enhanced oxidative stress in cells surrounding said senescent cells.

[0030] Aspect 24: The method of Aspect 1, wherein said SASP is associated with enhanced neutrophil activation in cells surrounding said senescent cells.

[0031] Aspect 25: The method of Aspect 1, wherein said SASP is associated with enhanced monocyte activation in cells surrounding said senescent cells.

[0032] Aspect 26: The method of Aspect 24, wherein said neutrophil activation is associated with release of superoxide radicals.

[0033] Aspect 27: The method of Aspect 24, wherein said neutrophil activation is associated with release of oxidative burst.

[0034] Aspect 28: The method of Aspect 24, wherein said neutrophil activation is associated with release of neutrophil extracellular traps.

[0035] Aspect 29: The method of Aspect 24, wherein said neutrophil activation is associated with enhanced antigen presentation.

[0036] Aspect 30: The method of Aspect 25, wherein said monocyte activation results in differentiation into macrophages.

[0037] Aspect 31 : The method of Aspect 25, wherein said monocyte activation involves enhanced expression of adhesion molecule ICAM-1.

[0038] Aspect 32: The method of Aspect 25, wherein said monocyte activation involves enhanced expression of adhesion molecule LFA-1.

[0039] Aspect 33: The method of Aspect 25, wherein said monocyte activation involves enhanced expression of adhesion molecule LFA-3.

[0040] Aspect 34: The method of Aspect 25, wherein said monocyte activation involves enhanced expression of adhesion molecule PEC AM-1.

[0041] Aspect 35: The method of Aspect 25, wherein said monocyte activation involves enhanced expression of adhesion molecule EpCAM.

[0042] Aspect 36: The method of Aspect 25, wherein said monocyte activation involves enhanced expression of adhesion molecule VLA4.

[0043] Aspect 37: The method of Aspect 25, wherein said monocyte activation results in differentiation into dendritic cells.

[0044] Aspect 38: The method of Aspect 25, wherein said monocyte activation is associated with enhanced T cell allostimulatory activity, inducing proliferation of allogeneic T cells.

[0045] Aspect 39: The method of Aspect 25, wherein said monocyte activation is associated with enhanced T cell allostimulatory activity, inducing cytokine production of allogeneic T cells.

[0046] Aspect 40: The method of Aspect 39, wherein said cytokine is IL-1 beta.

[0047] Aspect 41 : The method of Aspect 39, wherein said cytokine is IL-3.

[0048] Aspect 42: The method of Aspect 39, wherein said cytokine is IL-4.

[0049] Aspect 43: The method of Aspect 39, wherein said cytokine is IL-6.

[0050] Aspect 44: The method of Aspect 39, wherein said cytokine is IL-9.

[0051] Aspect 45: The method of Aspect 39, wherein said cytokine is IL-10.

[0052] Aspect 46: The method of Aspect 39, wherein said cytokine is IL-13.

[0053] Aspect 47: The method of Aspect 39, wherein said cytokine is IL-17.

[0054] Aspect 48: The method of Aspect 39, wherein said cytokine is IL-18.

[0055] Aspect 49: The method of Aspect 38, wherein said allostimulatory activity induces cytotoxic activity of allogeneic T cells, associated with expression of Granzyme B.

[0056] Aspect 50: The method of Aspect 38, wherein said allostimulatory activity induces cytotoxic activity of allogeneic T cells, associated with expression of perforin.

[0057] Aspect 51 : The method of Aspect 1, wherein said extracorporeal removal involves attaching ligands or antibodies specific to senescence-associated molecules to a solid matrix, and circulating blood or blood plasma over said matrix to remove said molecules.

[0058] Aspect 52: The method of Aspect 51, wherein said senescence-associated molecule is interleukin-6.

[0059] Aspect 53: The method of Aspect 51, wherein said senescence-associated molecule is HMGB 1.

[0060] Aspect 54: The method of Aspect 51, wherein said senescence-associated molecule is interleukin- 1 beta.

[0061] Aspect 55: The method of Aspect 51, wherein said senescence-associated molecule is senescence-associated beta-galactosidase.

[0062] Aspect 56: The method of Aspect 51, wherein said senescence-associated molecule is interleukin- 10 blocking protein.

[0063] Aspect 57: The method of Aspect 51, wherein said senescence-associated molecule is interleukin-8.

[0064] Aspect 58: The method of Aspect 51, wherein said senescence-associated molecule is interleukin-11.

[0065] Aspect 59: The method of Aspect 51, wherein said senescence-associated molecule is interleukin-12.

[0066] Aspect 60: The method of Aspect 51, wherein said senescence-associated molecule is interleukin-15.

[0067] Aspect 61 : The method of Aspect 51, wherein said senescence-associated molecule is interleukin- 17.

[0068] Aspect 62: The method of Aspect 51, wherein said senescence-associated molecule is interleukin- 17 A.

[0069] Aspect 63: The method of Aspect 51, wherein said senescence-associated molecule is interleukin- 17C.

[0070] Aspect 64: The method of Aspect 51, wherein said senescence-associated molecule is interleukin- 17F.

[0071] Aspect 65: The method of Aspect 51, wherein said senescence-associated molecule is interleukin- 18.

[0072] Aspect 66: The method of Aspect 51, wherein said senescence-associated molecule is interleukin-21.

[0073] Aspect 67: The method of Aspect 51, wherein said senescence-associated molecule is interleukin-22.

[0074] Aspect 68: The method of Aspect 51, wherein said senescence-associated molecule is interleukin-23.

[0075] Aspect 69: The method of Aspect 51, wherein said senescence-associated molecule is interleukin-27.

[0076] Aspect 70: The method of Aspect 51, wherein said senescence-associated molecule is interleukin-33.

[0077] Aspect 71 : The method of Aspect 51, wherein said senescence-associated molecule is VEGF.

[0078] Aspect 72: The method of Aspect 51, wherein said senescence-associated molecule is VEGF-A.

[0079] Aspect 73: The method of Aspect 51, wherein said senescence-associated molecule is VEGF-C.

[0080] Aspect 74: The method of Aspect 51, wherein said senescence-associated molecule is GDF-15.

[0081] Aspect 75: The method of Aspect 51, wherein said senescence-associated molecule is interferon gamma.

[0082] Aspect 76: The method of Aspect 51, wherein said senescence-associated molecule is IGFBP-2.

[0083] Aspect 77: The method of Aspect 51, wherein said senescence-associated molecule is IGFBP-5.

[0084] Aspect 78: The method of Aspect 51, wherein said senescence-associated molecule is IGFBP-7.

[0085] Aspect 79: The method of Aspect 51, wherein said senescence-associated molecule is MCP-1.

[0086] Aspect 80: The method of Aspect 51, wherein said senescence-associated molecule is TIMP-2.

[0087] Aspect 81 : The method of Aspect 51, wherein said senescence-associated molecule is TNF-alpha.

[0088] Aspect 82: The method of Aspect 51, wherein said senescence-associated molecule is lymphotoxin.

[0089] Aspect 83: The method of Aspect 1, wherein said senescence-associated factors are removed by binding to one or more antibodies specific to said senescence- associated factors.

[0090] Aspect 84: The method of Aspect 1, wherein said senescence-associated factors are removed by binding to one or more aptamers specific to said senescence- associated factors.

[0091] Aspect 85: The method of Aspect 1, wherein said senescence-associated factors are removed by binding to one or more proteins specific to said senescence- associated factors.

[0092] Aspect 86: The method of Aspect 1, wherein said senescence-associated factors are removed by binding to one or more microbodies specific to said senescence- associated factors.

[0093] Aspect 87: The method of Aspect 1, wherein said senescence-associated factors are removed by binding to one or more peptoids specific to said senescence- associated factors.

[0094] Aspect 88: The method of Aspect 1, wherein said senescence-associated factors are removed by binding to one or more peptides specific to said senescence- associated factors.

[0095] Aspect 89: A method of reducing effects of senescence in a subject, comprising: a) Administering one or more senolytic agents or therapies; b) Concurrently removing senescence-associated factors or extracellular vesicles; c) Optionally administering an anti-inflammatory mediator; and d) Optionally administering a regenerative cell or cell-free regenerative means, such as conditioned media.

[0096] Aspect 90: The method of Aspect 89, wherein said anti-inflammatory mediator is CNTF.

[0097] Aspect 91 : The method of Aspect 89, wherein said anti-inflammatory mediator is PGE-2.

[0098] Aspect 92: The method of Aspect 89, wherein said anti-inflammatory mediator is bcl-2.

[0099] Aspect 93: The method of Aspect 89, wherein said anti-inflammatory mediator is livin.

[0100] Aspect 94: The method of Aspect 89, wherein said anti-inflammatory mediator is survivin.

[0101] Aspect 95: The method of Aspect 89, wherein said anti-inflammatory mediator is membrane-bound TGF-beta.

[0102] Aspect 96: The method of Aspect 89, wherein said anti-inflammatory mediator is membrane-bound PD-L1.

[0103] Aspect 97: The method of Aspect 89, wherein said anti-inflammatory mediator is membrane-bound PD-L2.

[0104] Aspect 98: The method of Aspect 89, wherein said anti-inflammatory mediator is FoxP3.

[0105] Aspect 99: The method of Aspect 89, wherein said anti-inflammatory mediator is soluble PD-L1.

[0106] Aspect 100: The method of Aspect 89, wherein said anti-inflammatory mediator is soluble PD-L2.

[0107] Aspect 101 : The method of Aspect 89, wherein said senescent cell is inactivated by treatment with dasatinib.

[0108] Aspect 102: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with quercetin.

[0109] Aspect 103: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with dasatinib and quercetin.

[0110] Aspect 104: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with tetracycline.

[0111] Aspect 105: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with chlortetracycline.

[0112] Aspect 106: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with oxytetracycline.

[0113] Aspect 107: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with demeclocycline.

[0114] Aspect 108: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with methacycline.

[0115] Aspect 109: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with doxycycline.

[0116] Aspect 110: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with minocycline.

[0117] Aspect 111 : The method of Aspect 89, wherein said senescent cell is inactivated by treatment with tigecycline.

[0118] Aspect 112: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with pyrvinium.

[0119] Aspect 113: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with atovaquone.

[0120] Aspect 114: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with bedaquiline.

[0121] Aspect 115: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with irinotecan.

[0122] Aspect 116: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with sorafenib.

[0123] Aspect 117: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with niclosamide.

[0124] Aspect 118: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with stirpentol.

[0125] Aspect 119: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with chloroquine.

[0126] Aspect 120: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with rapamycin.

[0127] Aspect 121 : The method of Aspect 89, wherein said senescent cell is inactivated by treatment with a mitoriboscin.

[0128] Aspect 122: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with a mitoketoscin.

[0129] Aspect 123: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with a mitoflavoscin.

[0130] Aspect 124: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with 2-butene-l,4-bis-TPP.

[0131] Aspect 125: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with a derivative of 2-butene-l,4-bis-TPP.

[0132] Aspect 126: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with 2-chlorobenzyl-TPP.

[0133] Aspect 127: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with a derivative of 2-chlorobenzyl-TPP.

[0134] Aspect 128: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with 3-methylbenzyl-TPP.

[0135] Aspect 129: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with a derivative of 3-methylbenzyl-TPP.

[0136] Aspect 130: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with 2,4-dichlorobenzyl-TPP.

[0137] Aspect 131 : The method of Aspect 89, wherein said senescent cell is inactivated by treatment with a derivative of 2,4-dichlorobenzyl-TPP.

[0138] Aspect 132: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with 1 -naphthylmethyl -TPP.

[0139] Aspect 133: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with a derivative of 1-naphthylmethyl-TPP.

[0140] Aspect 134: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with p-xylylenebis-TPP.

[0141] Aspect 135: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with a derivative of p-xylylenebis-TPP.

[0142] Aspect 136: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with Vitamin C.

[0143] Aspect 137: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with fisetin.

[0144] Aspect 138: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with berberine.

[0145] Aspect 139: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with caffeic acid phenyl ester.

[0146] Aspect 140: The method of Aspect 89, wherein said senescent cell is inactivated by treatment with silibinin.

[0147] Aspect 141 : The method of Aspect 89, wherein said senescent cell is inactivated by treatment with brutieridin.

[0148] Aspect 142: The method of Aspect 89, wherein said regenerative cell is a pluripotent stem cell.

[0149] Aspect 143: The method of Aspect 142, wherein said pluripotent stem cell is an inducible pluripotent stem cell.

[0150] Aspect 144: The method of Aspect 142, wherein said pluripotent stem cell is a parthenogenic-derived pluripotent stem cell.

[0151] Aspect 145: The method of Aspect 142, wherein said pluripotent stem cell is a somatic cell nuclear transfer-derived pluripotent stem cell.

[0152] Aspect 146: The method of Aspect 142, wherein said pluripotent stem cell is a dedifferentiated somatic cell.

[0153] Aspect 147: The method of Aspect 89, wherein said regenerative cell is a tissue-specific progenitor cell capable of differentiating into endoderm, ectoderm, or mesoderm tissues, including pancreatic, liver, smooth muscle, striated muscle, cardiac muscle, bone, bone marrow, cartilage, spleen, thymus, lymph nodes, thyroid, epidermis, dermis, subcutaneous, heart, lung, vascular, endothelial, blood cells, bladder, kidney, digestive tract, esophagus, stomach, small intestine, large intestine, adipose, uterus, eye, testicular, ovarian, prostate, connective, endocrine, or mesentery tissue.

[0154] Aspect 148: The method of Aspect 146, wherein said dedifferentiated somatic cell is a dedifferentiated monocyte.

[0155] Aspect 149: The method of Aspect 148, wherein said monocyte is dedifferentiated by exposure to conditioned media from induced pluripotent stem cells (iPSCs).

[0156] Aspect 150: The method of Aspect 149, wherein said conditioned media is generated by culture of iPSC cells as embryoid bodies in a liquid media.

[0157] Aspect 151 : The method of Aspect 150, wherein said liquid media is Iscove’s media.

[0158] Aspect 152: The method of Aspect 150, wherein said liquid media is DMEM media.

[0159] Aspect 153: The method of Aspect 150, wherein said liquid media is OptiMEM media.

[0160] Aspect 154: The method of Aspect 150, wherein said liquid media is EMEM media.

[0161] Aspect 155: The method of Aspect 150, wherein said liquid media is RPMI- 1640 media.

[0162] Aspect 156: The method of Aspect 150, wherein said liquid media is AIM-V media.

[0163] Aspect 157: The method of Aspect 150, wherein said embryoid body is disassembled once every 2 days.

[0164] Aspect 158: The method of Aspect 150, wherein said embryoid body is disassembled once every 5 days.

[0165] Aspect 159: The method of Aspect 150, wherein said embryoid body is disassembled once every 10 days.

[0166] Aspect 160: The method of Aspect 150, wherein said embryoid body is composed of iPSC cells together with monocytes.

[0167] Aspect 161 : The method of Aspect 160, wherein said monocytes are first treated with a stressor before admixed with iPSCs for generation of conditioned media.

[0168] Aspect 162: The method of Aspect 161, wherein said stressor is hypoxia.

[0169] Aspect 163: The method of Aspect 161, wherein said stressor is hypertonicity.

[0170] Aspect 164: The method of Aspect 161, wherein said stressor is hypotonicity.

[0171] Aspect 165: The method of Aspect 161, wherein said stressor is hyperthermia.

[0172] Aspect 166: The method of Aspect 161, wherein said stressor is serum starvation.

[0173] Aspect 167: The method of Aspect 161, wherein said stressor is mTOR inhibition.

[0174] Aspect 168: The method of Aspect 161, wherein said stressor is AMPK activation.

[0175] Aspect 169: The method of Aspect 161, wherein said stressor is activation of inflammatory pathways.

[0176] Aspect 170: The method of Aspect 169, wherein said inflammatory pathway is MAP kinase.

[0177] Aspect 171 : The method of Aspect 169, wherein said inflammatory pathway is Janus Activated Kinase.

[0178] Aspect 172: The method of Aspect 169, wherein said inflammatory pathway is Signal Transducer and Activator of Transcription-3.

[0179] Aspect 173: The method of Aspect 169, wherein said inflammatory pathway is Signal Transducer and Activator of Transcription-5.

[0180] Aspect 174: The method of Aspect 169, wherein said inflammatory pathway is Signal Transducer and Activator of Transcription-6.

[0181] Aspect 175: The method of Aspect 169, wherein said inflammatory pathway is TLR2.

[0182] Aspect 176: The method of Aspect 175, wherein said TLR2 is activated by peptidoglycan.

[0183] Aspect 177: The method of Aspect 169, wherein said inflammatory pathway is TLR3.

[0184] Aspect 178: The method of Aspect 177, wherein said TLR3 is activated by double-stranded RNA.

[0185] Aspect 179: The method of Aspect 177, wherein said TLR3 is activated by Poly IC.

[0186] Aspect 180: The method of Aspect 177, wherein said TLR3 is activated by Poly LC.

[0187] Aspect 181 : The method of Aspect 177, wherein said TLR3 is activated by Poly IC:IC.

[0188] Aspect 182: The method of Aspect 177, wherein said TLR3 is activated by inactivated influenza virus.

[0189] Aspect 183: The method of Aspect 169, wherein said inflammatory pathway is TLR4.

[0190] Aspect 184: The method of Aspect 183, wherein said TLR4 activator is neutrophil extracellular traps.

[0191] Aspect 185: The method of Aspect 183, wherein said TLR4 activator is beta glucan.

[0192] Aspect 186: The method of Aspect 183, wherein said TLR4 activator is HMGB1.

[0193] Aspect 187: The method of Aspect 183, wherein said TLR4 activator is lipopolysaccharide.

[0194] Aspect 188: The method of Aspect 183, wherein said TLR4 activator is yeast cell wall extract.

[0195] Aspect 189: The method of Aspect 183, wherein said TLR4 activator is anti- TLR4 antibody.

[0196] Aspect 190: The method of Aspect 149, wherein said iPSC cells are engineered to express an inhibitor of TGF-beta.

[0197] Aspect 191 : The method of Aspect 190, wherein said inhibitor of TGF-beta is an antibody molecule.

[0198] Aspect 192: The method of Aspect 190, wherein said inhibitor of TGF-beta is a cameloid antibody.

[0199] Aspect 193: The method of Aspect 190, wherein said inhibitor of TGF-beta is a microbody.

[0200] Aspect 194: The method of Aspect 190, wherein said inhibitor of TGF-beta is an aptamer.

[0201] Aspect 195: The method of Aspect 190, wherein said inhibitor of TGF-beta is a molecule capable of inducing the process of RNA interference.

[0202] Aspect 196: The method of Aspect 195, wherein said molecule capable of inducing RNA interference is a short hairpin RNA.

[0203] Aspect 197: The method of Aspect 195, wherein said molecule capable of inducing RNA interference is a short interfering RNA.

[0204] Aspect 198: The method of Aspect 195, wherein said molecule capable of inducing RNA interference is a microRNA.

[0205] Aspect 199: The method of Aspect 195, wherein said molecule capable of inducing RNA interference is a long noncoding RNA.

[0206] Aspect 200: The method of Aspect 89, wherein said cell-free means is extracted or obtained from a regenerative cell, wherein said regenerative cell has been treated with a promoting agent that inhibits cellular differentiation while maintaining pluripotency.

[0207] Aspect 201 : The method of Aspect 200, wherein said promoting agent is GDF-11.

[0208] Aspect 202: The method of Aspect 200, wherein said promoting agent is GDF-15.

[0209] Aspect 203 : The method of Aspect 200, wherein said promoting agent is amniotic fluid.

[0210] Aspect 204: The method of Aspect 200, wherein said promoting agent is umbilical cord blood plasma.

[0211] Aspect 205: The method of Aspect 200, wherein said promoting agent is BMP2.

[0212] Aspect 206: The method of Aspect 200, wherein said promoting agent suppresses NF-kappa B activation.

[0213] Aspect 207: The method of Aspect 200, wherein said promoting agent increases NRF2 activation.

[0214] Aspect 208: The method of Aspect 200, wherein said promoting agent increases heme-oxygenase-1 activation.

[0215] Aspect 209: The method of Aspect 200, wherein said promoting agent increases bcl-2 activation.

[0216] Aspect 210: The method of Aspect 200, wherein said promoting agent increases bcl-2XL activation.

[0217] Aspect 211 : The method of Aspect 200, wherein said promoting agent increases survivin activation.

[0218] Aspect 212: The method of Aspect 200, wherein said promoting agent increases livin activation.

[0219] Aspect 213 : The method of Aspect 200, wherein said promoting agent is an HD AC inhibitor.

[0220] Aspect 214: The method of Aspect 213, wherein said HDAC inhibitor is valproic acid.

[0221] Aspect 215: The method of Aspect 213, wherein said HDAC inhibitor is trichostatin A.

[0222] Aspect 216: The method of Aspect 213, wherein said HDAC inhibitor is sodium phenylbutyrate.

[0223] Aspect 217: The method of Aspect 213, wherein said HDAC inhibitor is butyrate.

[0224] Aspect 218: The method of Aspect 200, wherein said promoting agent is a GSK-3 inhibitor.

[0225] Aspect 219: The method of Aspect 218, wherein said GSK-3 inhibitor is lithium.

[0226] Aspect 220: The method of Aspect 200, wherein said promoting agent causes a change in the genotype of the cell population.

[0227] Aspect 221 : The method of Aspect 200, wherein at least one promoting agent comprises or consists of an immortalizing oncogene.

[0228] Aspect 222: The method of Aspect 200, wherein said promoting agent is PIM I .

[0229] Aspect 223 : The method of Aspect 221, wherein said promoting agent is SV40 large T antigen.

[0230] Aspect 224: The method of Aspect 221, wherein said promoting agent is abll.

[0231] Aspect 225: The method of Aspect 221, wherein said promoting agent is AFF4.

[0232] Aspect 226: The method of Aspect 221, wherein said promoting agent is AKT2.

[0233] Aspect 227: The method of Aspect 221, wherein said promoting agent is AKL.

[0234] Aspect 228: The method of Aspect 221, wherein said promoting agent is AML I .

[0235] Aspect 229: The method of Aspect 221, wherein said promoting agent is MTG8.

[0236] Aspect 230: The method of Aspect 221, wherein said promoting agent is BCL6.

[0237] Aspect 231 : The method of Aspect 221, wherein said promoting agent is MCF2.

[0238] Aspect 232: The method of Aspect 221, wherein said promoting agent is DCF3.

[0239] Aspect 233 : The method of Aspect 221, wherein said promoting agent is EGFR.

[0240] Aspect 234: The method of Aspect 221, wherein said promoting agent is MLLT11.

[0241] Aspect 235: The method of Aspect 221, wherein said promoting agent is ERBB2.

[0242] Aspect 236: The method of Aspect 221, wherein said promoting agent is ETS1.

[0243] Aspect 237: The method of Aspect 221, wherein said promoting agent is CSF1R.

[0244] Aspect 238: The method of Aspect 221, wherein said promoting agent is FOS.

[0245] Aspect 239: The method of Aspect 221, wherein said promoting agent is FES.

[0246] Aspect 240: The method of Aspect 221, wherein said promoting agent is GN AS.

[0247] Aspect 241 : The method of Aspect 221, wherein said promoting agent is HER2.

[0248] Aspect 242: The method of Aspect 221, wherein said promoting agent is FGF3.

[0249] Aspect 243 : The method of Aspect 221, wherein said promoting agent is FGF4.

[0250] Aspect 244: The method of Aspect 221, wherein said promoting agent is JUN.

[0251] Aspect 245: The method of Aspect 221, wherein said promoting agent is c- kit.

[0252] Aspect 246: The method of Aspect 221, wherein said promoting agent is K- SAM.

[0253] Aspect 247: The method of Aspect 221, wherein said promoting agent is AKAP13.

[0254] Aspect 248: The method of Aspect 221, wherein said promoting agent is LCK.

[0255] Aspect 249: The method of Aspect 221, wherein said promoting agent is LM01.

[0256] Aspect 250: The method of Aspect 221, wherein said promoting agent is LYL I .

[0257] Aspect 251 : The method of Aspect 221, wherein said promoting agent is MASI.

[0258] Aspect 252: The method of Aspect 221, wherein said promoting agent is MDM2.

[0259] Aspect 253 : The method of Aspect 221, wherein said promoting agent is MOS.

[0260] Aspect 254: The method of Aspect 221, wherein said promoting agent is MYH11.

[0261] Aspect 255: The method of Aspect 221, wherein said promoting agent is MYB.

[0262] Aspect 256: The method of Aspect 221, wherein said promoting agent is MYCN.

[0263] Aspect 257: The method of Aspect 221, wherein said promoting agent is PAX5.

[0264] Aspect 258: The method of Aspect 221, wherein said promoting agent is RAF.

[0265] Aspect 259: The method of Aspect 221, wherein said promoting agent is RAS.

[0266] Aspect 260: The method of Aspect 221, wherein said promoting agent is REL.

[0267] Aspect 261 : The method of Aspect 221, wherein said promoting agent is ROSE

[0268] Aspect 262: The method of Aspect 221, wherein said promoting agent is SKI (PDGF-BB).

[0269] Aspect 263 : The method of Aspect 221, wherein said promoting agent is SET.

[0270] Aspect 264: The method of Aspect 221, wherein said promoting agent is SRC.

[0271] Aspect 265: The method of Aspect 221, wherein said promoting agent is TALI.

[0272] Aspect 266: The method of Aspect 221, wherein said promoting agent is TAN1.

[0273] Aspect 267: The method of Aspect 221, wherein said promoting agent is TIAN.

[0274] Aspect 268: The method of Aspect 221, wherein said promoting agent is TSC2.

[0275] Aspect 269: The method of Aspect 221, wherein said promoting agent is TRK.

[0276] Aspect 270: The method of Aspects 222 to 269, wherein said promoting agent can be conditionally inactivated.

[0277] Aspect 271 : The method of Aspect 147, wherein said tissue-specific progenitors are immortalized.

[0278] Aspect 272: The method of Aspect 271, wherein said immortalization is performed by transfection with an immortalizing factor.

[0279] Aspect 273 : The method of Aspect 272, wherein said immortalizing factor is one or more oncogenes selected from a group comprising: ABCB1, ABCG2, ABI1, ABL1, ABL2, ACKR3, ACSL3, ACSL6, ACVR1B, ACVR2A, AFF1, AFF3, AFF4, AKAP9, AKT1, AKT2, AKT3, ALDH1A1, ALDH2, ALK, AMER1, ANGPT1, ANGPT2, ANKRD23, APC, AR, ARAF, AREG, ARFRP1, ARHGAP26, ARHGEF12, ARID1A, ARID1B, ARID2, ARNT, ASPSCR1, ASXL1, ATF1, ATIC, ATM, ATP1A1, ATP2B3, ATR, ATRX, AURKA, AURKB, AXIN1, AXL, BAP1, BARD1, BBC3, BCL10, BCL11 A, BCL1 IB, BCL2, BCL2L1, BCL2L11, BCL2L2, BCL3, BCL6, BCL7A, BCL9, BCOR, BCORL1, BCR, BIRC3, BLM, BMPR1A, BRAF, BRCA1, BRCA2, BRD3, BRD4, BRINP3, BRIP1, BTG1, BTG2, BTK, BUB1B, Cl lorf30, C15orf65, C2orf44, CA6, CACNA1D, CALR, CAMTAI, CANT1, CARD11, CARS, CASC5, CASP8, CBFA2T3, CBFB, CBL, CBLB, CBLC, CCDC6, CCNB1IP1, CCND1, CCND2, CCND3, CCNE1, CD19, CD22, CD274, CD38, CD4, CD70, CD74, CD79A, CD79B, CD83, CDC73, CDH1, CDH11, CDK12, CDK4, CDK6, CDK7, CDK8, CDK9, CDKN1A, CDKN1B, CDKN2A, CDKN2B, CDKN2C, CDX2, CEBPA, CHCHD7, CHD2, CHD4, CHEK1, CHEK2, CHIC2, CHN1, CHORDCI, CIC, CIITA, CLP1, CLTC, CLTCL1, CNBP, CNOT3, CNTRL, COL1A1, COPB1, COX6C, CRBN, CREB1, CREB3L1, CREB3L2, CREBBP, CRKL, CRLF2, CRTC1, CRTC3, CSF1R, CSF3R, CTCF, CTLA4, CTNNA1, CTNNB1, CUL3, CXCR4, CYLD, CYP17A1, CYP2D6, DAXX, DDB2, DDIT3, DDR1, DDR2, DDX10, DDX3X, DDX5, DDX6, DEK, DICER1, DIS3, DLL4, DNM2, DNMT1, DNMT3A, DOT1L, DPYD, DUSP4, DUSP6, EBF1, ECT2L, EDNRB, EED, EGFR, EIF4A2, ELF4, ELK4, ELL, ELN, EML4, EP300, EPHA3, EPHA5, EPHA7, EPHA8, EPHB1, EPHB2, EPHB4, EPS15, ERBB2, ERBB3, ERBB4, ERC1, ERCC1, ERCC2, ERCC3, ERCC4, ERCC5, EREG, ERG, ERN1, ERRFI1, ESRI, ETV1, ETV4, ETV5, ETV6, EWSR1, EXT1, EXT2, EZH2, EZR, FAF1, FAIM3, FAM46C, FANCA, FANCC, FANCD2, FANCE, FANCF, FANCG, FANCL, FAS, FAT1, FBXO11, FBXW7, FCRL4, FEV, FGF10, FGF14, FGF19, FGF2, FGF23, FGF3, FGF4, FGF6, FGFR1, FGFR1OP, FGFR2, FGFR3, FGFR4, FH, FHIT, FIP1L1, FKBP1A, FLCN, FLU, FLT1, FLT3, FLT4, FNBP1, FOXA1, FOXL2, FOXO1, FOXO3, FOXO4, FOXP1, FRS2, FSTL3, FUBP1, FUS, GABRA6, GAS7, GATA1, GATA2, GATA3, GATA4, GATA6, GID4, GLI1, GMPS, GNA11, GNA12, GNA13,GNAQ, GNAS, GNRH1, G0LGA5, GOPC, GPC3, GPHN, GPR124, GRIN2A, GRAB, GSK3B, GUCY2C, H3F3A, H3F3B, HCK, HDAC1, HERPUD1, HEY1, HGF, HIP1, HIST1H1E, HIST1H3B, HIST1H4I, HLF, HMGA1, HMGA2, HMGN2P46, HNF1A, HNMT, HNRNPA2B1, HNRNPK, H00K3, H0XA11, H0XA13, H0XA9, HOXC11, HOXC13, HOXD11, HOXD13, HRAS, HSD3B1, HSP90AA1, HSP90AB1, IAPP, ID3, IDH1, IDH2, IGF1R, IGF2, IKBKE, IKZF1, IL2, IL21R, IL3RA, IL6, IL6ST, IL7R, INHBA, INPP4B, IRF2, IRF4, IRS2, ITGAV, ITGB1, ITK, ITPKB, JAK1, JAK2, JAK3, JAZF1, JUN, KAT6A, KAT6B, KCNJ5, KDM1A, KDM5A, KDM5C, KDM6A, KDR, KDSR, KEAP1, KEL, KIAA1549, KIF5B, KIR3DL1, KIT, KLF4, KLHL6, KLK2, KMT2A, KMT2C, KMT2D, KRAS, KTN1, LASPI, LCK, LCP1, LGALS3, LGR5, LHFP, LIFR, LM01, LM02, LOXL2, LPP, LRIG3, LRP1B, LUC7L2, LYL1, LYN, LZTR1, MAF, MAFB, MAGED1, MAGI2, MALT1, MAML2, MAP2K1, MAP2K2, MAP2K4, MAP3K1, MAPK1, MAPK11, MAX, MCL1, MDM2, MDM4, MDS2, MECOM, MED12, MEF2B, MEN1, MET, MITF, MKI67, MKL1, MLF1, MLH1, MLLT1, MLLT10, MLLT11, MLLT3, MLLT4, MLLT6, MMP9, MN1, MNX1, MPL, MRE1 1 A, MS4A1, MSH2, MSH6, MSI2, MSN, MST1R, MTCP1, MTF2, MTOR, MUC1, MUC16, MUTYH, MYB, MYC, MYCL, MYCN, MYD88, MYH11, MYH9, NACA, NAE1, NBN, NCKIPSD, NCOA1, NCOA2, NCOA4, NDRG1, NF1, NF2, NFE2L2, NFIB, NFKB2, NFKBIA, NIN, NKX2-1, NONO, NOTCH1, NOTCH2, NOTCH3, NPM1, NR4A3, NRAS, NSD1, NT5C2, NTRK1, NTRK2, NTRK3, NUMA1, NUP214, NUP93, NUP98, NUTM1, NUTM2B, OLIG2, OMD, P2RY8, PAFAH1B2, PAK3, PALB2, PARK2, PARP1, PATZ1, PAX3, PAX5, PAX7, PAX8, PBRM1, PBX1, PCM1, PCSK7, PDCD1, PDCD1LG2, PDE4DIP, PDGFB, PDGFRA, PDGFRB, PDK1, PECAM1, PERI, PHF6, PHOX2B, PICALM, PIK3C2B, PIK3CA, PIK3CB, PIK3CD, PIK3CG, PIK3R1, PIK3R2, PIM1, PLAG1, PLCG2, PML, PMS1, PMS2, POLDI, POLE, POTI, POU2AF1, POU5F1, PPARG, PPP2R1A, PRCC, PRDM1, PRDM16, PREX2, PRF1, PRKAR1A, PRKCI, PRKDC, PRLR, PRPF40B, PRRT2, PRRX1, PRSS8, PSIP1, PSMD4, PTBP1, PTCHI, PTEN, PTK2, PTPN11, PTPRC, PTPRD, QKI, RABEP1, RAC1, RAD21, RAD50, RAD51, RAD51B, RAD51C, RAD51D, RAFI, RALGDS, RANBP17, RANBP2, RAP1GDS1, RARA, R131, RBM10, RBM15, RCOR1, RECQL4, REL, RELN, RET, RHOA, RHOH, RICTOR, RIPK1, RMI2, RNF213, RNF43, ROS1, RPL10, RPL22, RPL5, RPN1, RPS6KB1, RPTOR, RUNX1, RUNX1T1, S1PR2, SAMHD1, SBDS, SDC4, SDHA, SDHAF2, SDHB, SDHC, SDHD, SEPT5, SEPT6, SEPT9, SET, SETBP1, SETD2, SF1, SF3A1, SF3B1, SF3B2, SFPQ, SGK1,SH2B3, SH3GL1, SLAMF7, SLC34A2, SLC45A3, SLIT2, SMAD2, SMAD3, SMAD4, SMARCA4, SMARCB1, SMARCE1, SMC1A, SMC3, SMO, SNCAIP, SNX29, SOCS1, SOXIO, SOX11, SOX2, SOX9, SPECC1, SPEN, SPOP, SPTA1, SRC, SRGAP3, SRSF2, SRSF3, SS18, SS18L1, SSX1, STAG2, STAT3, STAT4, STAT5B, STEAP1, STIL, STK11, SUFU, SUZ12, SYK, TAF1, TAF15, TALI, TAL2, TBL1XR1, TBX3, TCEA1, TCF12, TCF3, TCF7L2, TCL1A, TEK, TERC, TERT, TET1, TET2, TFE3, TFEB, TFG, TFPT, TFRC, TGFB1, TGFBR2, THRAP3, TIMP1, TJP1, TLX1, TLX3, TM7SF2, TMPRSS2, TNFAIP3, TNFRSF14, TNFRSF17, TNFRSF18, TNFRSF9, TNFSF11, TOPI, TOP2A, TP53, TP63, TPBG, TPM3, TPM4, TPR, TRAF2, TRAF3, TRAF3IP3, TRAF7, TRIM26, TRIM27, TRIM33, TRIP11, TRRAP, TSC1, TSC2, TSHR, TTK, TTL, TYMS, U2AF1, U2AF2, UBA1, UBR5, USP6, VEGFA, VEGFB, VHL, VPS51, VTI1A, WAS, WEE1, WHSCI, WHSC1L1, WIFI, WISP3, WNT11, WNT2B, WNT3, WNT3A, WNT4, WNT5A, WNT6, WNT7B, WRN, WT1, WWTR1, XBP1, XPA, XPC, XPO1, YWHAE, YWHAZ, ZAK, ZBTB16, ZBTB2, ZMYM2, ZMYM3, ZNF217, ZNF331, ZNF384, ZNF521, ZNF703, ZRSR2.

[0280] Aspect 274: A method for treating age-associated degeneration and / or stimulating regeneration in an organism, comprising administering one or more regenerative cells, together with one or more senolytic agents, while concurrently inducing dedifferentiation, wherein said dedifferentiation is induced by using retroviral expression means of dedifferentiation factors.

[0281] Aspect 275: The method of Aspect 274, wherein said retroviral expression vector is a pMXs vector.

[0282] Aspect 276: The method of Aspect 274, wherein said retroviral expression vector is a lentiviral vector.

[0283] Aspect 277: The method of Aspect 274, wherein said retroviral expression vector is an adenoviral vector.

[0284] Aspect 278: The method of Aspect 274, wherein said retroviral expression vector is a pMXs adeno-associated vector.

[0285] Aspect 279: The method of Aspect 274, wherein said retroviral expression vector is a herpesvirus vector.

[0286] Aspect 280: The method of Aspect 274, wherein said dedifferentiation genes are delivered by means of a plasmid-based vector.

[0287] Aspect 281 : The method of Aspect 274, wherein said dedifferentiation genes are delivered by means of an RNA-based vector.

[0288] Aspect 282: The method of Aspect 274, wherein said dedifferentiation genes are delivered by means of an mRNA-based vector.

[0289] Aspect 283 : The method of Aspect 274, wherein said dedifferentiation genes are delivered by means of a microRNA-based vector.

[0290] Aspect 284: The method of Aspect 274, wherein said dedifferentiation factors are administered together with ascorbic acid.

[0291] Aspect 285: The method of Aspect 274, wherein said dedifferentiation factors are administered together with a stimulator of the RAS pathway.

[0292] Aspect 286: The method of Aspect 274, wherein said dedifferentiation factors are administered together with a stimulator of the myc pathway.

[0293] Aspect 287: The method of Aspect 274, wherein said dedifferentiation factors are administered together with a stimulator of the PIM-1 pathway.

[0294] Aspect 288: The method of Aspect 274, wherein said dedifferentiation factors are administered together with a stimulator of the Janus activated kinase pathway.

[0295] Aspect 289: A method of reducing age-associated fibrosis in a patient, comprising administering T regulatory cells, and / or T regulatory cell-derived cell-free means together with one or more senolytic agents.

[0296] Aspect 290: The method of Aspect 289, wherein said T regulatory cells are generated subsequent to administration of mesenchymal stem cells or cell-free mesenchymal stem cell-derived means.

[0297] Aspect 291 : The method of Aspect 290, wherein said mesenchymal stem cells are selected for release of brain-derived neurotrophic factor upon stimulation with molecular signals associated with tissue injury.

[0298] Aspect 292: The method of Aspect 291, wherein mesenchymal stem cells are selected for expression of CD56.

[0299] Aspect 293: The method of Aspect 291, wherein mesenchymal stem cells produce at least 10 pg of brain-derived neurotrophic factor per milliliter subsequent to stimulation with Poly IC.

[0300] Aspect 294: The method of Aspect 291, wherein mesenchymal stem cells produce at least 25 pg of brain-derived neurotrophic factor per milliliter subsequent to stimulation with Poly IC.

[0301] Aspect 295: The method of Aspect 291, wherein mesenchymal stem cells produce at least 100 pg of brain-derived neurotrophic factor per milliliter subsequent to stimulation with Poly IC.

[0302] Aspect 296: The method of Aspect 290, wherein said mesenchymal stem cells are selected for release of ciliary neurotrophic growth factor upon stimulation with molecular signals associated with tissue injury.

[0303] Aspect 297: The method of Aspect 296, wherein mesenchymal stem cells are selected for expression of CD73.

[0304] Aspect 298: The method of Aspect 296, wherein mesenchymal stem cells produce at least 4 pg of ciliary neurotrophic growth factor per milliliter subsequent to stimulation with Poly IC.

[0305] Aspect 299: The method of Aspect 296, wherein mesenchymal stem cells produce at least 8 pg of ciliary neurotrophic growth factor per milliliter subsequent to stimulation with Poly IC.

[0306] Aspect 300: The method of Aspect 296, wherein mesenchymal stem cells produce at least 12 pg of ciliary neurotrophic growth factor per milliliter subsequent to stimulation with Poly IC.

[0307] Aspect 301 : The method of Aspect 290, wherein said mesenchymal stem cells are selected for release of basic fibroblast growth factor upon stimulation with molecular signals associated with tissue injury.

[0308] Aspect 302: The method of Aspect 301, wherein mesenchymal stem cells are selected for expression of CD 105.

[0309] Aspect 303: The method of Aspect 301, wherein mesenchymal stem cells produce at least 50 pg of basic fibroblast growth factor per milliliter subsequent to stimulation with Poly IC.

[0310] Aspect 304: The method of Aspect 301, wherein mesenchymal stem cells produce at least 100 pg of basic fibroblast growth factor per milliliter subsequent to stimulation with Poly IC.

[0311] Aspect 305: The method of Aspect 289, wherein said T regulatory cells and / or cell-free means secreted by said T regulatory cells are administered subsequent to administration of a fibrinolytic enzyme.

[0312] Aspect 306: The method of Aspect 305, wherein said enzyme is a matrix metalloprotease.

[0313] Aspect 307: The method of Aspect 306, wherein said matrix metalloprotease is MMP-3.

[0314] Aspect 308: The method of Aspect 306, wherein said matrix metalloprotease is MMP-5.

[0315] Aspect 309: The method of Aspect 306, wherein said matrix metalloprotease is MMP-6.

[0316] Aspect 310: The method of Aspect 306, wherein said matrix metalloprotease is MMP-9.

[0317] Aspect 311 : The method of Aspect 306, wherein said matrix metalloprotease is MMP-12.

[0318] Aspect 312: The method of Aspect 289, wherein said T regulatory cells and / or cell-free means generated by said T regulatory cells are administered subsequent to administration of an inhibitor of NOGO.

[0319] Aspect 313: The method of Aspect 312, wherein said inhibitor of NOGO is an antisense oligonucleotide.

[0320] Aspect 314: The method of Aspect 312, wherein said inhibitor of NOGO is one or more molecules capable of inducing RNA interference. Aspect 315: The method of Aspect 314, wherein said molecule capable of inducing RNA interference is a short interfering RNA.

[0321] Aspect 316: The method of Aspect 314, wherein said molecule capable of inducing RNA interference is a short hairpin RNA.

[0322] Aspect 317: The method of Aspect 312, wherein said inhibitor of NOGO is a small molecule inhibitor.

[0323] Aspect 318: The method of Aspect 312, wherein said inhibitor of NOGO is an antibody.

[0324] Aspect 319: The method of Aspect 312, wherein said inhibitor of NOGO is an aptamer.

[0325] Aspect 320: The method of Aspect 312, wherein said inhibitor of NOGO is a somamer.

[0326] Aspect 321 : The method of Aspect 312, wherein said inhibitor of NOGO is a bispecific antibody.

[0327] Aspect 322: The method of Aspect 312, wherein said inhibitor of NOGO is a ribozyme.

[0328] Aspect 323 : The method of Aspect 312, wherein said inhibitor of NOGO is a microantibody.

[0329] Aspect 324: The method of Aspect 312, wherein said inhibitor of NOGO is a hammerhead ribozyme.

[0330] Aspect 325: The method of Aspect 312, wherein said inhibitor of NOGO is a soluble receptor.

[0331] Aspect 326: The method of Aspect 289, wherein said T regulatory cells are administered subsequent to administration of a myeloid lineage cell.

[0332] Aspect 327: The method of Aspect 326, wherein said myeloid lineage cell is a myeloid-derived dendritic cell.

[0333] Aspect 328: The method of Aspect 327, wherein said myeloid dendritic cell is immature.

[0334] Aspect 329: The method of Aspect 328, wherein said myeloid dendritic cell is generated from a monocytic progenitor.

[0335] Aspect 330: The method of Aspect 329, wherein said monocyte is isolated from peripheral blood.

[0336] Aspect 331 : The method of Aspect 329, wherein said monocyte is isolated from peripheral blood that is mobilized by treatment with G-CSF.

[0337] Aspect 332: The method of Aspect 329, wherein said monocyte is isolated from peripheral blood that is mobilized by treatment with GM-CSF.

[0338] Aspect 333: The method of Aspect 329, wherein said monocyte is isolated from peripheral blood that is mobilized by treatment with M-CSF.

[0339] Aspect 334: The method of Aspect 329, wherein said monocyte is isolated from peripheral blood that is mobilized by treatment with interleukin-3.

[0340] Aspect 335: The method of Aspect 329, wherein said monocyte is isolated from peripheral blood that is mobilized by treatment with interleukin-10.

[0341] Aspect 336: The method of Aspect 329, wherein said monocyte is isolated from peripheral blood that is mobilized by treatment with beta glucan.

[0342] Aspect 337: The method of Aspect 329, wherein said monocyte is isolated from peripheral blood that is mobilized by treatment with interferon gamma.

[0343] Aspect 338: The method of Aspect 329, wherein said monocyte is isolated from menstrual blood.

[0344] Aspect 339: The method of Aspect 329, wherein said monocyte is isolated from umbilical cord blood.

[0345] Aspect 340: The method of Aspect 329, wherein said monocyte is isolated from Wharton’s Jelly.

[0346] Aspect 341 : The method of Aspect 329, wherein said monocyte is isolated from bone marrow.

[0347] Aspect 342: The method of Aspect 329, wherein said monocyte is isolated from adipose tissue.

[0348] Aspect 343: The method of Aspect 329, wherein said monocyte is isolated from omental tissue.

[0349] Aspect 344: The method of Aspect 329, wherein said monocyte is pretreated with one or more agents to induce a tolerogenic phenotype prior to differentiation into a myeloid dendritic cell with immature phenotype, and exosomes are derived from said myeloid dendritic cells.

[0350] Aspect 345: The method of Aspect 344, wherein said monocyte is pretreated with PGE2 at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0351] Aspect 346: The method of Aspect 344, wherein said monocyte is pretreated with PGE2 at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0352] Aspect 347: The method of Aspect 344, wherein said monocyte is pretreated with PGE2 at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0353] Aspect 348: The method of Aspect 344, wherein said monocyte is pretreated with genistein at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0354] Aspect 349: The method of Aspect 344, wherein said monocyte is pretreated with genistein at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0355] Aspect 350: The method of Aspect 344, wherein said monocyte is pretreated with genistein at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0356] Aspect 351 : The method of Aspect 344, wherein said monocyte is pretreated with quercetin at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0357] Aspect 352: The method of Aspect 344, wherein said monocyte is pretreated with quercetin at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0358] Aspect 353: The method of Aspect 344, wherein said monocyte is pretreated with quercetin at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0359] Aspect 354: The method of Aspect 344, wherein said monocyte is pretreated with hypertonic saline at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0360] Aspect 355: The method of Aspect 344, wherein said monocyte is pretreated with hypertonic saline at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0361] Aspect 356: The method of Aspect 344, wherein said monocyte is pretreated with hypertonic saline at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0362] Aspect 357: The method of Aspect 344, wherein said monocyte is pretreated with VEGF at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0363] Aspect 358: The method of Aspect 344, wherein said monocyte is pretreated with VEGF at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0364] Aspect 359: The method of Aspect 344, wherein said monocyte is pretreated with VEGF at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0365] Aspect 360: The method of Aspect 344, wherein said monocyte is pretreated with PDGF-BB at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0366] Aspect 361 : The method of Aspect 344, wherein said monocyte is pretreated with PDGF-BB at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0367] Aspect 362: The method of Aspect 344, wherein said monocyte is pretreated with PDGF-BB at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0368] Aspect 363: The method of Aspect 344, wherein said monocyte is pretreated with IGF-1 at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0369] Aspect 364: The method of Aspect 344, wherein said monocyte is pretreated with IGF-1 at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0370] Aspect 365: The method of Aspect 344, wherein said monocyte is pretreated with IGF-1 at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0371] Aspect 366: The method of Aspect 344, wherein said monocyte is pretreated with salinomycin at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0372] Aspect 367: The method of Aspect 344, wherein said monocyte is pretreated with salinomycin at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0373] Aspect 368: The method of Aspect 344, wherein said monocyte is pretreated with salinomycin at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0374] Aspect 369: The method of Aspect 344, wherein said monocyte is pretreated with erythropoietin at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0375] Aspect 370: The method of Aspect 344, wherein said monocyte is pretreated with erythropoietin at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0376] Aspect 371 : The method of Aspect 344, wherein said monocyte is pretreated with erythropoietin at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0377] Aspect 372: The method of Aspect 344, wherein said monocyte is pretreated with NF-kappa B decoy oligonucleotides at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0378] Aspect 373: The method of Aspect 344, wherein said monocyte is pretreated with NF-kappa B decoy oligonucleotides at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0379] Aspect 374: The method of Aspect 344, wherein said monocyte is pretreated with NF-kappa B decoy oligonucleotides at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0380] Aspect 375: The method of Aspect 344, wherein said monocyte is pretreated with Ikk-B decoy oligonucleotides at a sufficient concentration and for a time period tostimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0381] Aspect 376: The method of Aspect 344, wherein said monocyte is pretreated with Ikk-B decoy oligonucleotides at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0382] Aspect 377: The method of Aspect 344, wherein said monocyte is pretreated with Ikk-B decoy oligonucleotides at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0383] Aspect 378: The method of Aspect 344, wherein said monocyte is pretreated with interleukin-35 at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0384] Aspect 379: The method of Aspect 344, wherein said monocyte is pretreated with interleukin-35 at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0385] Aspect 380: The method of Aspect 344, wherein said monocyte is pretreated with interleukin-35 at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0386] Aspect 381 : The method of Aspect 344, wherein said monocyte is pretreated with TGF-beta at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0387] Aspect 382: The method of Aspect 344, wherein said monocyte is pretreated with TGF-beta at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0388] Aspect 383: The method of Aspect 344, wherein said monocyte is pretreated with TGF-beta at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0389] Aspect 384: The method of Aspect 344, wherein said monocyte is pretreated with ascorbic acid at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0390] Aspect 385: The method of Aspect 344, wherein said monocyte is pretreated with ascorbic acid at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0391] Aspect 386: The method of Aspect 344, wherein said monocyte is pretreated with ascorbic acid at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0392] Aspect 387: The method of Aspect 344, wherein said monocyte is pretreated with n-acetylcysteine at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0393] Aspect 388: The method of Aspect 344, wherein said monocyte is pretreated with n-acetylcysteine at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0394] Aspect 389: The method of Aspect 344, wherein said monocyte is pretreated with n-acetylcysteine at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0395] Aspect 390: The method of Aspect 344, wherein said monocyte is pretreated with alpha lipoic acid at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes. Aspect

[0396] 391 : The method of Aspect 344, wherein said monocyte is pretreated with alpha lipoic acid at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0397] Aspect 392: The method of Aspect 344, wherein said monocyte is pretreated with alpha lipoic acid at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0398] Aspect 393: The method of Aspect 344, wherein said monocyte is pretreated with methylene blue at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0399] Aspect 394: The method of Aspect 344, wherein said monocyte is pretreated with methylene blue at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0400] Aspect 395: The method of Aspect 344, wherein said monocyte is pretreated with methylene blue at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0401] Aspect 396: The method of Aspect 344, wherein said monocyte is pretreated with valproic acid at a sufficient concentration and for a time period to stimulate production of at least 10 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0402] Aspect 397: The method of Aspect 344, wherein said monocyte is pretreated with valproic acid at a sufficient concentration and for a time period to stimulate production of at least 20 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0403] Aspect 398: The method of Aspect 344, wherein said monocyte is pretreated with valproic acid at a sufficient concentration and for a time period to stimulate production of at least 40 pg / ml of interleukin- 10 from a culture of 1 million monocytes.

[0404] Aspect 399: The method of Aspect 344, wherein said immature dendritic cell expresses interleukin- 10.

[0405] Aspect 400: The method of Aspect 344, wherein said immature dendritic cell expresses interleukin-4. Aspect 401 : The method of Aspect 344, wherein said immature dendritic cell expresses interleukin-13.

[0406] Aspect 402: The method of Aspect 344, wherein said immature dendritic cell expresses interleukin-20. A

[0407] spect 403: The method of Aspect 344, wherein said immature dendritic cell expresses TGF-beta.

[0408] Aspect 404: The method of Aspect 344, wherein said immature dendritic cell expresses interleukin-22.

[0409] Aspect 405: The method of Aspect 344, wherein said immature dendritic cell expresses interleukin-35.

[0410] Aspect 406: The method of Aspect 344, wherein said immature dendritic cell expresses interleukin-37.

[0411] Aspect 407: The method of Aspect 344, wherein said immature dendritic cell expresses interleukin-38.

[0412] Aspect 408: The method of Aspect 344, wherein said immature dendritic cell expresses endoglin.

[0413] Aspect 409: The method of Aspect 344, wherein said immature dendritic cell expresses VEGF.

[0414] Aspect 410: The method of Aspect 344, wherein said immature dendritic cell expresses HLA-G.

[0415] Aspect 411 : The method of Aspect 344, wherein said immature dendritic cell expresses interleukin- 12 p40 homodimer.

[0416] Aspect 412: The method of Aspect 344, wherein said immature dendritic cell possesses increased phagocytic activity as compared to a mature dendritic cell.

[0417] Aspect 413: The method of Aspect 344, wherein said immature dendritic cell possesses increased migratory activity towards chemotactic gradients as compared to a mature dendritic cell.

[0418] Aspect 414: The method of Aspect 413, wherein said chemotactic gradient is comprised of the chemokine IL-8.

[0419] Aspect 415: The method of Aspect 413, wherein said chemotactic gradient is comprised of the chemokine MIP-1 alpha.

[0420] Aspect 416: The method of Aspect 413, wherein said chemotactic gradient is comprised of the chemokine MIP-1 beta. A

[0421] spect 417: The method of Aspect 413, wherein said chemotactic gradient is comprised of the chemokine CXCL12.

[0422] Aspect 418: The method of Aspect 413, wherein said chemotactic gradient is comprised of the chemokine MCP-1.

[0423] Aspect 419: The method of Aspect 413, wherein said chemotactic gradient is comprised of the chemokine TNF-alpha.

[0424] Aspect 420: The method of Aspect 413, wherein said chemotactic gradient is comprised of the chemokine lymphotoxin.

[0425] Aspect 421 : The method of Aspect 413, wherein said chemotactic gradient is comprised of the chemokine hyaluronic acid degradation products.

[0426] Aspect 422: The method of Aspect 413, wherein said chemotactic gradient is comprised of the chemokine HMGB1.

[0427] Aspect 423: The method of Aspect 344, wherein said immature dendritic cell possesses decreased HL A II compared to a mature dendritic cell.

[0428] Aspect 424: The method of Aspect 344, wherein said immature dendritic cell possesses decreased HL A I compared to a mature dendritic cell.

[0429] Aspect 425: The method of Aspect 344, wherein said immature dendritic cell possesses decreased CD la compared to a mature dendritic cell.

[0430] Aspect 426: The method of Aspect 344, wherein said immature dendritic cell possesses decreased CD40 compared to a mature dendritic cell.

[0431] Aspect 427: The method of Aspect 344, wherein said immature dendritic cell possesses decreased CD80 compared to a mature dendritic cell.

[0432] Aspect 428: The method of Aspect 344, wherein said immature dendritic cell possesses decreased CD86 compared to a mature dendritic cell.

[0433] Aspect 429: The method of Aspect 344, wherein said immature dendritic cell possesses decreased interleukin- 15 receptor compared to a mature dendritic cell.

[0434] Aspect 430: The method of Aspect 344, wherein said immature dendritic cell possesses decreased interferon gamma compared to a mature dendritic cell.

[0435] Aspect 431 : The method of Aspect 344, wherein said immature dendritic cell possesses decreased interleukin- 18 receptor compared to a mature dendritic cell.

[0436] Aspect 432: The method of Aspect 344, wherein said immature dendritic cell possesses decreased progesterone receptor compared to a mature dendritic cell.

[0437] Aspect 433: The method of Aspect 344, wherein said immature dendritic cell possesses decreased c-kit compared to a mature dendritic cell.

[0438] Aspect 434: The method of Aspect 344, wherein said immature dendritic cell is capable of inducing generation of T regulatory cells upon culture with allogeneic naive T cells.

[0439] Aspect 435: The method of Aspect 434, wherein said immature dendritic cells upregulate expression of AIRE upon contact with said allogeneic naive T cells.

[0440] Aspect 436: The method of Aspect 435, wherein said dendritic cells upregulate expression of AIRE in an interleukin- 10 dependent manner.

[0441] Aspect 437: The method of Aspect 435, wherein said dendritic cells upregulate expression of AIRE in a TGF-beta dependent manner.

[0442] Aspect 438: The method of Aspect 435, wherein said dendritic cells upregulate expression of AIRE in a soluble HLA-G dependent manner.

[0443] Aspect 439: The method of Aspect 435, wherein said dendritic cells upregulate expression of AIRE in an endoglin dependent manner.

[0444] Aspect 440: The method of Aspect 435, wherein said dendritic cells upregulate expression of AIRE in an FGF-1 dependent manner.

[0445] Aspect 441 : The method of Aspect 435, wherein said dendritic cells upregulate expression of AIRE in an FGF-2 dependent manner.

[0446] Aspect 442: The method of Aspect 435, wherein said dendritic cells upregulate expression of AIRE in an FGF-5 dependent manner.

[0447] Aspect 443: The method of Aspect 435, wherein said dendritic cells upregulate expression of AIRE in a NOTCH dependent manner.

[0448] Aspect 444: The method of Aspect 326, wherein said myeloid lineage cell is an immature neutrophil.

[0449] Aspect 445: The method of Aspect 444, wherein said immature neutrophil is a neutrophil progenitor.

[0450] Aspect 446: The method of Aspect 444, wherein said immature neutrophil is capable of differentiating along the neutrophilic lineage or the monocytic lineage.

[0451] Aspect 447: The method of Aspect 444, wherein said immature neutrophil expresses PU1.1.

[0452] Aspect 448: The method of Aspect 444, wherein said immature neutrophil expresses G-CSF receptor.

[0453] Aspect 449: The method of Aspect 444, wherein said immature neutrophil expresses stem cell factor receptor.

[0454] Aspect 450: The method of Aspect 444, wherein said immature neutrophil expresses c-met.

[0455] Aspect 451 : The method of Aspect 444, wherein said immature neutrophil expresses M-CSF receptor.

[0456] Aspect 452: The method of Aspect 444, wherein said immature neutrophil expresses GM-CSF receptor.

[0457] Aspect 453 : The method of Aspect 444, wherein said immature neutrophil produces interleukin- 10 upon stimulation with a TLR-4 agonist.

[0458] Aspect 454: The method of Aspect 453, wherein said TLR-4 agonist is beta glucan.

[0459] Aspect 455: The method of Aspect 453, wherein said TLR-4 agonist is hyaluronic acid degradation products.

[0460] Aspect 456: The method of Aspect 453, wherein said TLR-4 agonist is HMGB1.

[0461] Aspect 457: The method of Aspect 453, wherein said TLR-4 agonist is histones.

[0462] Aspect 458: The method of Aspect 326, wherein said myeloid lineage cell is a myeloid suppressor cell.

[0463] Aspect 459: The method of Aspect 458, wherein said myeloid suppressor cell is capable of producing Reptimed.

[0464] Aspect 460: The method of Aspect 458, wherein said myeloid suppressor cell is capable of differentiating into monocytes when treated with All Trans Retinoic Acid.

[0465] Aspect 461 : The method of Aspect 458, wherein said myeloid suppressor cell is capable of differentiating into monocytes when treated with vitamin D3.

[0466] Aspect 462: The method of Aspect 458, wherein said myeloid suppressor cell is capable of differentiating into monocytes when treated with M-CSF.

[0467] Aspect 463: The method of Aspect 458, wherein said myeloid suppressor cell is capable of differentiating into monocytes when treated with GM-CSF.

[0468] Aspect 464: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of nitric oxide.

[0469] Aspect 465: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of reactive oxygen species.

[0470] Aspect 466: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of superoxide.

[0471] Aspect 467: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of hydrogen peroxide

[0472] .Aspect 468: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of arginase.

[0473] Aspect 469: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of interleukin- 10.

[0474] Aspect 470: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of soluble PD-L1.

[0475] Aspect 471 : The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of soluble VISTA.

[0476] Aspect 472: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of LAG-3.

[0477] Aspect 473: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of TIM3.

[0478] Aspect 474: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of PGE2.

[0479] Aspect 475: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of secreted vimentin.

[0480] Aspect 476: The method of Aspect 458, wherein said myeloid suppressor cell inhibits T cell proliferation by production of secreted calreticulin.

[0481] Aspect 477: The method of Aspect 289, wherein said T regulatory cell is administered together with a mesenchymal cell differentiated to the oligodendrocyte lineage.

[0482] Aspect 478: The method of Aspect 289, wherein said T regulatory cell is administered together with a pluripotent stem cell differentiated to the oligodendrocyte lineage.

[0483] Aspect 479: The method of Aspect 478, wherein said pluripotent stem cell is generated by transfecting a somatic cell with cytoplasm from an oocyte.

[0484] Aspect 480: The method of Aspect 479, wherein said cytoplasm is transfected by use of electroporation.

[0485] Aspect 481 : The method of Aspect 479, wherein said cytoplasm is transfected by use of cell fusion.

[0486] Aspect 482: The method of Aspect 479, wherein said cytoplasm is transfected by use of streptolysin O to generate transient holes in the cytoplasm of said recipient cell.

[0487] Aspect 483 : The method of Aspect 479, wherein said cytoplasm is transfected by use of cell penetrating peptides to generate transient holes in the cytoplasm of said recipient cell.

[0488] Aspect 484: The method of Aspect 478, wherein said pluripotent stem cell is an induced pluripotent stem cell.

[0489] Aspect 485: The method of Aspect 484, wherein said induced pluripotent stem cell is created by transfection of cells with pluripotency inducing factors.

[0490] Aspect 486: The method of Aspect 485, wherein said pluripotency inducing factor is OCT4.

[0491] Aspect 487: The method of Aspect 485, wherein said pluripotency inducing factor is PIM-1.

[0492] Aspect 488: The method of Aspect 485, wherein said pluripotency inducing factor is NANOG.

[0493] Aspect 489: The method of Aspect 485, wherein said pluripotency inducing factor is c-met.

[0494] Aspect 490: The method of Aspect 485, wherein said pluripotency inducing factor is hTERT.

[0495] Aspect 491 : The method of Aspect 485, wherein said pluripotency inducing factor is KLF4.

[0496] Aspect 492: The method of Aspect 485, wherein said pluripotency inducing factor is RAS.

[0497] Aspect 493 : The method of Aspect 485, wherein said pluripotency inducing factor is NOTCH.

[0498] Aspect 494: The method of Aspect 485, wherein said pluripotency inducing factor is BMP2.

[0499] Aspect 495: The method of Aspect 485, wherein said pluripotency inducing factor is BMP4.

[0500] Aspect 496: The method of Aspect 485, wherein said pluripotency inducing factor is AIRE.

[0501] Aspect 497: A method of regenerating aged tissue, comprising administering T regulatory cell-derived cell-free means, wherein said T regulatory cells are optionally treated with one or more histone deacetylase inhibitors.

[0502] Aspect 498: The method of Aspect 497, wherein said cell-free means are exosomes.

[0503] Aspect 499: The method of Aspect 498, wherein said cell-free means are exosomes expressing CD8.

[0504] Aspect 500: The method of Aspect 498, wherein said cell-free means are exosomes expressing phosphatidylserine.

[0505] Aspect 501 : The method of Aspect 498, wherein said cell-free means are apoptotic bodies.

[0506] Aspect 502: The method of Aspect 498, wherein said cell-free means are ferroptotic bodies.

[0507] Aspect 503: The method of Aspect 498, wherein said cell-free means is conditioned media.

[0508] Aspect 504: The method of Aspect 498, wherein said cell-free means is conditioned media from stressed cells.

[0509] Aspect 505: The method of Aspect 504, wherein said cellular stress is exposure to inflammatory stimuli.

[0510] Aspect 506: The method of Aspect 505, wherein said inflammatory stimuli is activation of one or more danger-associated receptors.

[0511] Aspect 507: The method of Aspect 506, wherein said danger-associated receptor is a toll -like receptor.

[0512] Aspect 508: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with beta glucan.

[0513] Aspect 509: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with Poly IC.

[0514] Aspect 510: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with Poly LC.

[0515] Aspect 511 : The method of Aspect 507, wherein said toll-like receptor is activated by treatment with CpG DNA.

[0516] Aspect 512: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with BCG.

[0517] Aspect 513: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with flagellin.

[0518] Aspect 514: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with lipopolysaccharide.

[0519] Aspect 515: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with low molecular weight hyaluronic acid.

[0520] Aspect 516: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with free histone.

[0521] Aspect 517: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with Isoxazolo[5,4-d]pyrimidine.

[0522] Aspect 518: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with imiquimod.

[0523] Aspect 519: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with Poly A:U.

[0524] Aspect 520: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with MPL.

[0525] Aspect 521 : The method of Aspect 507, wherein said toll-like receptor is activated by treatment with Poly G3.

[0526] Aspect 522: The method of Aspect 507, wherein said toll-like receptor is activated by treatment with Poly G10.

[0527] Aspect 523 : The method of Aspect 507, wherein said toll-like receptor is activated by treatment with hyperthermia-exposed cells.

[0528] Aspect 524: The method of Aspect 523, wherein said hyperthermia-exposed cells possess an upregulation of hsp90 of more than 50% as compared to untreated cells of the same type.

[0529] Aspect 525: The method of Aspect 497, wherein a histone deacetylase inhibitor is administered locally and / or systemically to enhance reparative activity of said T regulatory cell.

[0530] Aspect 526: The method of Aspect 525, wherein said histone deacetylase inhibitor i s CXD 101.

[0531] Aspect 527: The method of Aspect 525, wherein said histone deacetylase inhibitor i s HD AC 10-IN - 1.

[0532] Aspect 528: The method of Aspect 525, wherein said histone deacetylase inhibitor is Tubastatin A TFA.

[0533] Aspect 529: The method of Aspect 525, wherein said histone deacetylase inhibitor is ACY-775.

[0534] Aspect 530: The method of Aspect 525, wherein said histone deacetylase inhibitor is Panobinostat.

[0535] Aspect 531 : The method of Aspect 525, wherein said histone deacetylase inhibitor is Trichostatin A.

[0536] Aspect 532: The method of Aspect 525, wherein said histone deacetylase inhibitor is Vorinostat.

[0537] Aspect 533: The method of Aspect 525, wherein said histone deacetylase inhibitor is Entinostat.

[0538] Aspect 534: The method of Aspect 525, wherein said histone deacetylase inhibitor is BML-210.

[0539] Aspect 535: The method of Aspect 525, wherein said histone deacetylase inhibitor is Abexinostat.

[0540] Aspect 536: The method of Aspect 525, wherein said histone deacetylase inhibitor is Dacinostat.

[0541] Aspect 537: The method of Aspect 525, wherein said histone deacetylase inhibitor is Quisinostat.

[0542] Aspect 538: The method of Aspect 525, wherein said histone deacetylase inhibitor is Mocetinostat.

[0543] Aspect 539: The method of Aspect 525, wherein said histone deacetylase inhibitor is Valproic Acid.

[0544] Aspect 540: The method of Aspect 525, wherein said histone deacetylase inhibitor is CUDC-101.

[0545] Aspect 541 : The method of Aspect 525, wherein said histone deacetylase inhibitor is GSK3117391.

[0546] Aspect 542: The method of Aspect 525, wherein said histone deacetylase inhibitor is Droxinostat.

[0547] Aspect 543: The method of Aspect 525, wherein said histone deacetylase inhibitor is MCI 568.

[0548] Aspect 544: The method of Aspect 525, wherein said histone deacetylase inhibitor is Pracinostat. Aspect

[0549] 545: The method of Aspect 525, wherein said histone deacetylase inhibitor isDivalproex Sodium.

[0550] Aspect 546: The method of Aspect 525, wherein said histone deacetylase inhibitor is Diferuloylmethane.

[0551] Aspect 547: The method of Aspect 525, wherein said histone deacetylase inhibitor is Sodium butyrate.

[0552] Aspect 548: The method of Aspect 525, wherein said histone deacetylase inhibitor is PCI-34051.

[0553] Aspect 549: The method of Aspect 525, wherein said histone deacetylase inhibitor is SR-4370.

[0554] Aspect 550: The method of Aspect 525, wherein said histone deacetylase inhibitor is Givinostat.

[0555] Aspect 551 : The method of Aspect 525, wherein said histone deacetylase inhibitor is Tubacin.

[0556] Aspect 552: The method of Aspect 525, wherein said histone deacetylase inhibitor is AR-42.

[0557] Aspect 553: The method of Aspect 525, wherein said histone deacetylase inhibitor is (-)-Parthenolide.

[0558] Aspect 554: The method of Aspect 525, wherein said histone deacetylase inhibitor is Tubastatin A HC1.

[0559] Aspect 555: The method of Aspect 525, wherein said histone deacetylase inhibitor is Resminostat.

[0560] Aspect 556: The method of Aspect 525, wherein said histone deacetylase inhibitor is CUDC-907.

[0561] Aspect 557: The method of Aspect 525, wherein said histone deacetylase inhibitor is M344.

[0562] Aspect 558: The method of Aspect 525, wherein said histone deacetylase inhibitor is Tacedinaline.

[0563] Aspect 559: The method of Aspect 525, wherein said histone deacetylase inhibitor is Romidepsin.

[0564] Aspect 560: The method of Aspect 525, wherein said histone deacetylase inhibitor is 4-Phenylbutyric acid.

[0565] Aspect 561 : The method of Aspect 525, wherein said histone deacetylase inhibitor is Sinapinic acid.

[0566] Aspect 562: The method of Aspect 525, wherein said histone deacetylase inhibitor is Sulforaphane.

[0567] Aspect 563: The method of Aspect 525, wherein said histone deacetylase inhibitor is UFO 10.

[0568] Aspect 564: The method of Aspect 525, wherein said histone deacetylase inhibitor is Suberohydroxamic acid.

[0569] Aspect 565: The method of Aspect 525, wherein said histone deacetylase inhibitor is NKL 22.

[0570] Aspect 566: The method of Aspect 525, wherein said histone deacetylase inhibitor is ITSA-1.

[0571] Aspect 567: The method of Aspect 525, wherein said histone deacetylase inhibitor is KA2507.

[0572] Aspect 568: The method of Aspect 525, wherein said histone deacetylase inhibitor is Isoguanosine.

[0573] Aspect 569: The method of Aspect 525, wherein said histone deacetylase inhibitor is Raddeanin A.

[0574] Aspect 570: The method of Aspect 525, wherein said histone deacetylase inhibitor is BRD3308.

[0575] Aspect 571 : The method of Aspect 525, wherein said histone deacetylase inhibitor is TH34.

[0576] Aspect 572: The method of Aspect 525, wherein said histone deacetylase inhibitor is Tinostamustine.

[0577] Aspect 573: The method of Aspect 497, wherein exosomes from a pluripotent stem cell-derived mesenchymal stem cell population are administered systemically from either a culture vessel or through the use of a bioreactor.

[0578] Aspect 574: The method of Aspect 573, wherein said mesenchymal stem cell is transfected with a growth factor.

[0579] Aspect 575: The method of Aspect 574, wherein said growth factor is HGF- 1.

[0580] Aspect 576: The method of Aspect 574, wherein said growth factor is FGF- 1.

[0581] Aspect 577: The method of Aspect 574, wherein said growth factor is EGF- 1.

[0582] Aspect 578: The method of Aspect 574, wherein said growth factor is angiopoietin.

[0583] Aspect 579: The method of Aspect 574, wherein said growth factor is placental growth factor.

[0584] Aspect 580: The method of Aspect 574, wherein said growth factor is adiponectin.

[0585] Aspect 581 : The method of Aspect 574, wherein said growth factor is vasoactive intestinal peptide precursor.

[0586] Aspect 582: The method of Aspect 574, wherein said growth factor is endoglin.

[0587] Aspect 583 : The method of Aspect 574, wherein said growth factor is myostatin.

[0588] Aspect 584: The method of Aspect 574, wherein said growth factor is TGF- beta.

[0589] Aspect 585: The method of Aspect 574, wherein said growth factor is vascular endothelial growth factor.

[0590] Aspect 586: The method of Aspect 574, wherein said growth factor is GDF- 11.

[0591] Aspect 587: The method of Aspect 574, wherein said growth factor is GDF- 15.

[0592] Aspect 588: The method of Aspect 574, wherein said growth factor is hyaluronic acid synthase.

[0593] Aspect 589: The method of Aspect 574, wherein said growth factor is interleukin-33.

[0594] Aspect 590: The method of Aspect 574, wherein said growth factor is osteosarcoma-derived growth factor.

[0595] Aspect 591 : The method of Aspect 574, wherein said growth factor is midkine.

[0596] Aspect 592: The method of Aspect 574, wherein said growth factor is PDGF-BB.

[0597] Aspect 593 : The method of Aspect 574, wherein said growth factor is IGF-1.

[0598] Aspect 594: The method of Aspect 574, wherein said growth factor is nerve growth factor.

[0599] Aspect 595: A method of enhancing efficacy of regenerative medicine interventions, comprising extracorporeally removing senescence-associated secretory factors.

[0600] Aspect 596: The method of Aspect 595, wherein said extracorporeal removal of senescence-associated secretory factors and / or extracellular vesicles is performed by contacting whole blood, plasma, or serum from said selected patient with a substrate that is specific for one or more senescence-associated molecules.

[0601] Aspect 597: The method of Aspect 596, wherein said substrate is an antibody.

[0602] Aspect 598: The method of Aspect 597, wherein said antibody is a human IgM, IgG, IgA, IgD, or IgE antibody.

[0603] Aspect 599: The method of Aspect 597, wherein said antibody is a lama or camel antibody.

[0604] Aspect 600: The method of Aspect 597, wherein said antibody is a shark antibody.

[0605] Aspect 601 : The method of Aspect 596, wherein said substrate is a microbody.

[0606] Aspect 602: The method of Aspect 596, wherein said substrate is an aptamer.

[0607] Aspect 603: The method of Aspect 596, wherein said substrate is a peptide. Aspect 604: The method of Aspect 596, wherein said substrate is a peptoid.

[0608] Aspect 605: The method of Aspect 596, wherein said substrate is a quantum dot.

[0609] Aspect 606: The method of Aspect 597, wherein said antibody is specific for a non-polymorphic region of MHC I.

[0610] Aspect 607: The method of Aspect 596, wherein said substrate comprises a silica gel.

[0611] Aspect 608: The method of Aspect 596, wherein said substrate comprises dextran.

[0612] Aspect 609: The method of Aspect 596, wherein said substrate comprises agarose.

[0613] Aspect 610: The method of Aspect 596, wherein said substrate comprises a nylon polymer.

[0614] Aspect 611 : The method of Aspect 596, wherein said substrate comprises an acrylic acid polymer.

[0615] Aspect 612: The method of Aspect 596, wherein said substrate comprises a co-polymer of ethylene and maleic acid anhydride.

[0616] Aspect 613: The method of Aspect 596, wherein said substrate comprises aminopropylsilica.

[0617] Aspect 614: The method of Aspect 596, wherein said substrate comprises aminocelite.

[0618] Aspect 615: The method of Aspect 596, wherein said substrate comprises glass beads.

[0619] Aspect 616: The method of Aspect 596, wherein said substrate comprises silicate-containing diatomaceous earth.

[0620] Aspect 617: The method of Aspect 596, wherein said substrate is a filter membrane.

[0621] Aspect 618: The method of Aspect 596, wherein said substrate comprises a plurality of filter membranes arranged in sequence.

[0622] Aspect 619: The method of Aspect 596, further comprising returning contacted whole blood, plasma, or serum to the patient, wherein said contacted whole blood, plasma, or serum contains substantially fewer senescence-associated factors and / or extracellular vesicles in comparison to whole blood, plasma, or serum originally residing in the patient.

[0623] Aspect 620: The method of Aspect 596, wherein said substrate comprises a porous exterior of a hollow fiber filter.

[0624] Aspect 621 : The method of Aspect 596, wherein the antibody that is specific for senescence-associated antigen is immobilized on the porous exterior of the hollowfiber filter and wherein the hollow fiber filter is configured to allow passage of blood cells through the interior of said hollow fiber filter and allow diffusion of senescence- associated factors to the porous exterior.

[0625] Aspect 622: The method of Aspect 596, wherein said senescence-associated factor is interleukin- 11.

[0626] Aspect 623: The method of Aspect 596, wherein said senescence-associated factor is interleukin- 1.

[0627] Aspect 624: The method of Aspect 596, wherein said senescence-associated factor is interleukin- 1 beta.

[0628] Aspect 625: The method of Aspect 596, wherein said senescence-associated factor is interleukin-2 binding protein or interleukin-2 soluble receptor.

[0629] Aspect 626: The method of Aspect 596, wherein said senescence-associated factor is interleukin-6.

[0630] Aspect 627: The method of Aspect 596, wherein said senescence-associated factor is interleukin-7.

[0631] Aspect 628: The method of Aspect 596, wherein said senescence-associated factor is interleukin-9.

[0632] Aspect 629: The method of Aspect 596, wherein said senescence-associated factor is interleukin-12.

[0633] Aspect 630: The method of Aspect 596, wherein said senescence-associated factor is interleukin-13.

[0634] Aspect 631 : The method of Aspect 596, wherein said senescence-associated factor is interleukin-15.

[0635] Aspect 632: The method of Aspect 596, wherein said senescence-associated factor is interleukin- 17.

[0636] Aspect 633: The method of Aspect 596, wherein said senescence-associated factor is interleukin- 17 A.

[0637] Aspect 634: The method of Aspect 596, wherein said senescence-associated factor is interleukin- 17C.

[0638] Aspect 635: The method of Aspect 596, wherein said senescence-associated factor is interleukin- 17F.

[0639] Aspect 636: The method of Aspect 596, wherein said senescence-associated factor is interleukin- 18.

[0640] Aspect 637: The method of Aspect 596, wherein said senescence-associated factor is interleukin-21.

[0641] Aspect 638: The method of Aspect 596, wherein said senescence-associated factor is interleukin-22.

[0642] Aspect 639: The method of Aspect 596, wherein said senescence-associated factor is interleukin-23.

[0643] Aspect 640: The method of Aspect 596, wherein said senescence-associated factor is interleukin-27.

[0644] Aspect 641 : The method of Aspect 596, wherein said senescence-associated factor is interleukin-33.

[0645] Aspect 642: The method of Aspect 596, wherein said senescence-associated factor is TRAIL.

[0646] Aspect 643: The method of Aspect 596, wherein said senescence-associated factor is BLyS.

[0647] Aspect 644: The method of Aspect 596, wherein said senescence-associated factor is LIGHT.

[0648] Aspect 645: The method of Aspect 596, wherein said senescence-associated factor is TRANCE.

[0649] Aspect 646: The method of Aspect 596, wherein said senescence-associated factor is HMGB1.

[0650] Aspect 647: The method of Aspect 596, wherein said senescence-associated factor is lymphotoxin.

[0651] Aspect 648: The method of Aspect 596, wherein said senescence-associated factor is drug resistance-related protein LRP.

[0652] Aspect 649: The method of Aspect 596, wherein said senescence-associated factor is major vault protein.

[0653] Aspect 650: The method of Aspect 596, wherein said senescence-associated factor is thyroid hormone binding protein precursor.

[0654] Aspect 651 : The method of Aspect 596, wherein said senescence-associated factor is prolyl 4-hydroxylase, beta subunit precursor.

[0655] Aspect 652: The method of Aspect 596, wherein said senescence-associated factor is chain A, human protein disulfide isomerase.

[0656] Aspect 653: The method of Aspect 596, wherein said senescence-associated factor is electron-transfer-flavoprotein, beta polypeptide.

[0657] Aspect 654: The method of Aspect 596, wherein said senescence-associated factor is ATP synthase, H+ transporting, mitochondrial Fl complex, alpha subunit.

[0658] Aspect 655: The method of Aspect 596, wherein said senescence-associated factor is cathepsin B.

[0659] Aspect 656: The method of Aspect 596, wherein said senescence-associated factor is Ig gamma-2B chain C region.

[0660] Aspect 657: The method of Aspect 596, wherein said senescence-associated factor is haptoglobin.

[0661] Aspect 658: The method of Aspect 596, wherein said senescence-associated factor is IGF -BP.

[0662] Aspect 659: The method of Aspect 596, wherein said senescence-associated factor is free histones.

[0663] Aspect 660: The method of Aspect 596, wherein said senescence-associated factor is Ig gamma-2A chain C region secreted form.

[0664] Aspect 661 : The method of Aspect 596, wherein said senescence-associated factor is Ig kappa chain C region.

[0665] Aspect 662: The method of Aspect 596, wherein said senescence-associated factor is Ig mu chain C region.

[0666] Aspect 663: The method of Aspect 596, wherein said senescence-associated factor is chemokine subfamily B Cys-X-Cys.

[0667] Aspect 664: The method of Aspect 596, wherein said senescence-associated factor is platelet glycoprotein lb alpha chain.

[0668] Aspect 665: The method of Aspect 596, wherein said senescence-associated factor is cathepsin S.

[0669] Aspect 665: The method of Aspect 596, wherein said senescence-associated factor is inter alpha-trypsin inhibitor, heavy chain 4.

[0670] Aspect 667: The method of Aspect 596, wherein said senescence-associated factor is arachidonate 12-lipoxygenase, 12S-type.

[0671] Aspect 668: The method of Aspect 596, wherein said senescence-associated factor is immunoglobulin kappa variable 5-39.

[0672] Aspect 669: The method of Aspect 596, wherein said senescence-associated factor is indolethylamine N-methyltransferase.

[0673] Aspect 670: The method of Aspect 596, wherein said senescence-associated factor is kinesin-like protein; kinesin-like protein KIF21B.

[0674] Aspect 671 : The method of Aspect 596, wherein said senescence-associated factor is platelet glycoprotein lb beta chain.

[0675] Aspect 672: The method of Aspect 596, wherein said senescence-associated factor is plastin-1.

[0676] Aspect 673: The method of Aspect 595, wherein said regenerative intervention is administration of a regenerative cell.

[0677] Aspect 674: The method of Aspect 673, wherein said regenerative cell is a stem cell.

[0678] Aspect 675: The method of Aspect 674, wherein said stem cell is a hematopoietic stem cell.

[0679] Aspect 676: The method of Aspect 675, wherein said hematopoietic stem cell possesses the ability to generate lymphoid, myeloid, erythroid, or megakaryocytic cells.

[0680] Aspect 677: The method of Aspect 675, wherein said hematopoietic stem cell expresses the marker CD34.

[0681] Aspect 678: The method of Aspect 675, wherein said hematopoietic stem cell expresses the marker CD 133.

[0682] Aspect 679: The method of Aspect 675, wherein said hematopoietic stem cell expresses the marker CD 105.

[0683] Aspect 680: The method of Aspect 675, wherein said hematopoietic stem cell expresses the marker IL-3 receptor.

[0684] Aspect 681 : The method of Aspect 675, wherein said hematopoietic stem cell expresses the marker IL-7 receptor.

[0685] Aspect 682: The method of Aspect 675, wherein said hematopoietic stem cell expresses the marker c-mpl.

[0686] Aspect 683: The method of Aspect 675, wherein said hematopoietic stem cell expresses the marker Fas ligand.

[0687] Aspect 684: The method of Aspect 675, wherein said hematopoietic stem cell proliferates in response to blockade of TGF-beta signaling.

[0688] Aspect 685: The method of Aspect 595, wherein said regenerative medicine intervention is administration of very small embryonic-like stem cells.

[0689] Aspect 686: The method of Aspect 595, wherein said regenerative medicine intervention is administration of regenerative cell conditioned media.

[0690] Aspect 687: The method of Aspect 595, wherein said regenerative medicine intervention is administration of mesenchymal stem cells.

[0691] Aspect 688: The method of Aspect 687, wherein said mesenchymal stem cell expresses IL- 10 receptor.

[0692] Aspect 689: The method of Aspect 687, wherein said mesenchymal stem cell expresses HLA-G.

[0693] Aspect 690: The method of Aspect 687, wherein said mesenchymal stem cell expresses c-kit.

[0694] Aspect 691 : The method of Aspect 687, wherein said mesenchymal stem cell expresses PD-L1.

[0695] Aspect 692: The method of Aspect 687, wherein said mesenchymal stem cell expresses c-met.

[0696] Aspect 693 : The method of Aspect 687, wherein said mesenchymal stem cell expresses IL-22 receptor.DETAILED DESCRIPTION

[0697] The detailed description set forth below is intended as a description of various configurations of the subject technology and is not intended to represent the only configurations in which the subject technology may be practiced. The detailed description includes specific details for the purpose of providing an understanding of the subject technology. It will be apparent to those skilled in the art that the subject technology may be practiced without these specific details.

[0698] In accordance with the disclosed invention, provided are methods of stimulation of regeneration and / or augmentation of immunity and / or stimulation of angiogenesis using extracorporeal techniques to remove senescence associated factors and / or extracellular vesicles from circulation.

[0699] By “administration” or “administering” is meant a method of providing a dosage of an agent of the invention to a mammal (e.g., a human), where the route is, e.g., topical, oral, parenteral (e.g., intravenous, intraperitoneal, intrarterial, intradermal, intramuscular, or subcutaneous injection, inhalation, optical drops, or implant), nasal, vaginal, rectal, or sublingual application in admixture with a pharmaceutically acceptable carrier adapted for such use. The preferred method of administration can vary depending on various factors, e.g., the components of the pharmaceutical composition, site of the potential or actual disease (e.g., the location of lung, breast, colon, prostate, liver, brain, heart, etc.), and the severity of disease.

[0700] By “analog” is meant an agent that differs from, but is structurally, functionally, and / or chemically related to the reference agent. The analog may retain the essential properties, functions, or structures of the reference agent. Most preferably, the analog retains at least one biological function of the reference agent. Generally, differences are limited so that the structure or sequence of the reference agent and the analog are similar overall. For example, a peptide analog and its reference peptide may differ in amino acid sequence by one or more amino acid substitutions, additions, and / or deletions, or the presence of one or more non-naturally occurring amino acid residues, in any combination. An analog of a peptide or polypeptide of the invention may be naturally occurring, such as an allelic variant, or it may be a variant that is not known to occur naturally. Non-naturally occurring analogs of peptides may be made by direct synthesis, by modification, or by mutagenesis techniques.

[0701] By “chelating agent” is meant a molecule that forms multiple chemical bonds with a single metal atom. Prior to forming the bonds, the chelating agent has more than one pair of unshared electrons. The bonds are formed by sharing pairs of electrons with the metal atom. Chelating agents include, for example, an iminodicarboxylic group or a polyaminopolycarboxylic group. Chelating agents may be attached to an agent of the invention using the methods generally described in Liu et al., Bioconjugate Chem. 12(4):653, 2001; Alter et al., U.S. Pat. No. 5,753,627; and PCT Publication No. WO 91 / 01144; each of which is hereby incorporated by reference). An agent of the invention may be complexed, through its attached chelating agent, to a detectable label, thereby resulting in an agent that is indirectly labeled. Similarly, cytotoxic or therapeutic agents, may also be attached via a chelating group to an agent of the invention.

[0702] By “coupled” is meant the characteristic of a first molecule being joined to a second molecule by a covalent bond or through noncovalent intermolecular attraction.

[0703] By “cytotoxic agent” is meant any naturally occurring, modified, or synthetic compound that is toxic to cells. Such agents are useful in the treatment of neoplasms, and in the treatment of other symptoms or diseases characterized by cell proliferation or a hyperactive cell population. Cytotoxic agents can also be used to target undesirable cells or tissues other than neoplastic cells or tissues, e.g., senescent cells. Cytotoxic agents include, but are not limited to, alkylating agents, antibiotics, antimetabolites, tubulin inhibitors, topoisomerase I and II inhibitors, hormonal agonists or antagonists, immunomodulators, or agents that cause cell lysis including naturally occurring or synthetic peptides. Cytotoxic agents may be cytotoxic when activated by light or infrared (Photofrin, IR dyes; Nat. Biotechnol. 19(4):327-331, 2001), may operate through other mechanistic pathways, or be supplementary potentiating agents.

[0704] By “detectable label” is meant any type of label which, when attached to an agent of the invention, renders the agent detectable. A detectable label may be toxic or non-toxic, and may have one or more of the following attributes, without restriction: fluorescence (Kiefer et al., W0 9740055), color, toxicity (e.g., radioactivity, e.g., a y- emitting radionuclide, Auger-emitting radionuclide, P-emitting radionuclide, an a- emitting radionuclide, or a positron-emitting radionuclide), radiosensitivity, or photosensitivity. Although a detectable label may be directly attached, for example, to an amino acid residue of an agent of the invention, or indirectly attached, for example, by being complexed with a chelating group that is attached (e.g., linked via a covalent bond or indirectly linked) to an amino acid residue of an agent of the invention. A detectable label may also be indirectly attached to an agent of the invention by the ability of the label to be specifically bound by a second molecule. One example of this type of an indirectly attached label is a biotin label that can be specifically bound by a second molecule, streptavidin. The second molecule may also be linked to a moiety that allows neutron capture (e.g., a boron cage as described in, for example, Kahl et al., Proc. Natl. Acad. Sci. USA 87:7265-7269, 1990).

[0705] A detectable label may also be a metal ion from heavy elements or rare earth ions, such as Gd3+, Fe3+, Mn3+, or Cr2+ (see, e.g., Curter, Invest. Radiol. 33(10):752- 761, 1998). Preferred radioactive detectable labels are radioactive iodine labels (e.g.,1221, 1231, 1241, 1251, or 1311) that are capable of being coupled to each D- or L-Tyr or D- or L-4-amino-Phe residues present in the agents of the invention. Preferred nonradioactive detectable labels are the many known dyes that are capable of being coupled to NH2 -terminal amino acid residues.

[0706] Preferred examples of detectable labels that may be toxic to cells include ricin, diphtheria toxin, and radioactive detectable labels (e.g., 1221, 1231, 1241, 1251, 1311, 177Lu, 64Cu, 67Cu, 153Sm, 166Ho, 186Re, 188Re, 211 At, 212Bi, 225Ac, 67Ga, 68Ga, 75Br, 76Br, 77Br, 117mSn, 47Sc, 109Pd, 89Sr, 159Gd, 49Pm, 142Pr, l l lAg, Dy, 213Bi, U lin, 114mln, 201Ti 195mPt, 193Pt, 86Y and 90Y). These compounds, and others described herein may be directly or indirectly attached to an agent of the invention or its analogs. A toxic detectable label may also be a chemotherapeutic agent (e.g., camptothecins, homocamptothecins, 5 -fluorouracil or adriamycin), or may be a radiosensitizing agent (e.g., paclitaxel, gemcitabine, fluoropyrimidine, metronitozil, or the deoxycytidine analog 2', 2' difluoro-2'-deoxycytidine (dFdCyd) to which is directly or indirectly attached an agent or analog thereof of the present invention.

[0707] A detectable label, when coupled to an agent of the invention emits a signal that can be detected by a signal transducing machine. In some cases, the detectable label can emit a signal spontaneously, such as when the detectable label is a radionuclide. In other cases the detectable label emits a signal as a result of being stimulated by an external field such as when the detectable label is a relaxivity metal. Examples of signals include, without limitation, gamma rays, X-rays, visible light, infrared energy, and radio waves. Examples of signal transducing machines include, without limitation, gamma cameras including SPECT / CT devices, PET scanners, fluorimeters, and Magnetic Resonance Imaging (MRI) machines.

[0708] By “diagnostically effective amount” is meant a dose of detectably-labeled agent that, when administered internally to a mammal, is quantitatively sufficient to be detected by a signal transducing machine external to the mammal (e.g., a gamma camera used in gamma scintigraphy) but typically is quantitatively insufficient to produce a pharmacological effect.

[0709] By “imaging agent” is meant a compound that, when administered to a living subject, such as a mammal (e.g., a human), allows the visualization of internal structures(e.g., cells, tissues, and organs) and, in some cases can provide information as to the function of a cell, tissue, or organ in the subject.

[0710] By “linker moiety” is meant a sequence of amino acid residues, e.g., at least one, two, three, four, five, six, seven, eight, nine, ten, fifteen, twenty, thirty, forty, fifty, or more residues, that couples an agent of the invention (e.g., a peptide, polypeptide, protein, small molecule, antibody, or antibody fragment that target senescent cells) to, e.g., one or more of a detectable label, a therapeutic agent, and a chelating agent.

[0711] By “senescent cell” is meant a cell that is metabolically active but is permanently withdrawn from the cell cycle (see, e.g., Campisi, Cell 120:513-522, 2005). Senescent cells do not replicate and possess one or more of the following additional characteristics attributed to senescent cells: cell cycle arrest in the G1 phase; an enlarged, flattened morphology; increased granularity; staining for P-galactosidase activity at pH 6; senescence associated heterochromatic foci; and characteristic gene expression that is in part regulated by pl6 and p21. Alternatively, a senescent cell is a cell that can be induced to become senescent (e.g., by stress) or that expresses cell-surface markers characteristic of senescent cells; these markers include senescent cell-specific antigens having the polypeptide sequences set forth in SEQ ID NOS: 11-23. Senesecent cell-specific antigens are those peptides, polypeptides, or glycoproteins that are expressed on the cell surface of senescent cells, but are absent or only weakly expressed on the cell surface of nonsenescent cells.

[0712] By “peptide” is meant a polymer that includes two or more amino acids joined to each other by a peptide bond or a modified peptide bond. “Peptide” refers to both short chain polymers, commonly referred to as peptides, oligopeptides, or oligomers, having, e.g., about 10-50 linked amino acid residues, and to longer polymers having up to about 100 amino acid residues in length. Peptides may contain amino acids other than the 20 gene-encoded amino acids, and linkages other than peptide bonds and may include cyclic or branched peptides. “Peptides” include amino acid sequences modified either by natural processes, or by chemical modification techniques that are well known in the art. Modifications may occur anywhere in a polypeptide, including the peptide backbone, the amino acid side-chains, and the amino or carboxyl termini.

[0713] The notations used herein for the peptide amino acid residues are those abbreviations commonly used in the art. The less common abbreviations Abu, Ava, [3- Ala, hSer, Nle, Nva, Pal, Dab, and Dap stand for 2-amino-butyric acid, amino valeric acid, beta-aminopropionic acid, homoserine, norleucine, norvaline, (2, 3, or 4) 3-pyridyl- Ala, 1,4-diaminobutyric acid, and 1,3 -diaminopropionic acid, respectively. In all aspects of the invention, it is noted that when amino acids are not designated as either D- or L- amino acids, the amino acid is either an L-amino acid or could be either a D- or L-amino acid. Examples of peptides of the invention include those peptides having the sequence of, or a sequence substantially identical to, the sequences set forth in SEQ ID NOs: 1-3 and 5-8 and related sequences. These peptide sequences can also be incorporated into a polypeptide, protein, antibody, or antibody fragment.

[0714] By “reducing agent” is meant a chemical compound used to reduce another chemical compound by donating electrons, thereby becoming oxidized.

[0715] By “specifically binds” is meant that an agent of the invention recognizes and binds to a target (e.g., a senescent cell), but does not substantially recognize and bind to a non-target (e.g., non-senescent cells), both in vivo and in a sample, e.g., an in vitro biological sample, that includes, e.g., senescent cells. A desirable agent of the invention specifically binds to senescent cells. Preferably, the agents of the invention bind senescent cells with at least 2, 5, 10, 20, 100, or 1000 fold greater affinity than they bind to nonsenescent cells. Alternatively, agents of the invention specifically bind to senescent cells with a dissociation constant less than 10-6M, more preferably less than 10-7M, 10-8M, 10-9M, 10-10M, 10-1 IM, or 10-12M, and most preferably less than 10-13M, 10-14M, or 10-15M.

[0716] By “substantial sequence identity” or “substantially identical” is meant that a nucleic acid or amino acid sequence exhibits at least 50%, preferably 60%, 70%, 75%, or 80%, more preferably 85%, 90% or 95%, and most preferably 99% identity to a reference amino acid sequence (e.g., one or more of the sequences set forth in SEQ ID NOs: 1-3 and 5-8). For amino acid sequences, the length of comparison sequences will generally be at least 5 amino acids, preferably at least 10 contiguous amino acids, more preferably at least 15, 20, 25, 30, 40, 50, 60, 80, 90, 100, 150, 200, 250, 300, or 350 contiguous amino acids, and most preferably the full-length amino acid sequence. Sequence identity is typically measured using BLAST® (Basic Local Alignment Search Tool) or BLAST®2with the default parameters specified therein (see, Altschul et al., J. Mol. Biol. 215:403- 410, 1990); and Tatiana et al., FEMS Microbiol. Lett. 174:247-250, 1999). This software program matches similar sequences by assigning degrees of homology to various substitutions, deletions, and other modifications. Conservative substitutions typically include substitutions within the following groups: glycine, alanine, valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine.

[0717] By “therapeutic agent” is meant any compound that is used in the detection, diagnosis or treatment of disease. Such compounds may be naturally-occurring, modified, or synthetic. A therapeutic agent may be, for example, an agent that causes apoptosis or necrosis of a cell (e.g., a senescent cell) in an organism (e.g., a mammal, such as a human), thereby reducing the number of such cells in the organism. Therapeutic agents that reduce the number of senescent cells in an organism may be, e.g., alkylating agents, antibiotics, antimetabolites, hormonal agonists or antagonists, anti- or pro-apoptotic agents, immunomodulators, or supplementary potentiating agents.

[0718] By “treating, stabilizing, or preventing cancer” is meant causing a reduction in the size of a tumor or in the number of cancer cells, slowing or preventing an increase in the size of a tumor or in cancer cell proliferation, increasing the disease-free survival time between the disappearance of a tumor or other cancer and its reappearance, preventing an initial or subsequent occurrence of a tumor or other cancer, or reducing an adverse symptom associated with a tumor or other cancer. In a desired embodiment, the percent of tumor or cancerous cells surviving the treatment is at least 20, 40, 60, 80, or 100% lower than the initial number of tumor or cancerous cells, as measured using any standard assay, such as those described herein. Desirably, the decrease in the number of tumor or cancerous cells induced by administration of a compound of the invention is at least 2, 5, 10, 20, or 50-fold greater than the decrease in the number of non-tumor or non- cancerous cells. Desirably, the methods of the present invention result in a decrease of 20, 40, 60, 80, or 100% in the size of a tumor or number of cancerous cells as determined using standard methods. Desirably, at least 20, 40, 60, 80, 90, or 95% of the treated subjects have a complete remission in which all evidence of the tumor or cancer disappears. Desirably, the tumor or cancer does not reappear after more than 5, 10, 15, or 20 years.

[0719] By “treating, stabilizing, or preventing age-related diseases” is meant causing a reduction in the number of symptoms, a decrease in severity of any, all, or substantially all of the symptoms, a complete resolution of any, all, or substantially all symptoms, or preventing the occurrence of any, all, or substantially all symptoms associated with one or more of the age-related diseases including, but not limited to, cardiovascular disease, cerebrovascular disease, peripheral vascular disease, Alzheimer's disease, osteoarthritis, cardiac diastolic dysfunction, benign prostatic hypertrophy, and cancers that increase in incidence and prevalence with increasing patient age such as cancer, e.g., breast cancer, prostate cancer, and colon cancer.

[0720] By “treating, stabilizing, or preventing tobacco-related diseases” is meant causing a reduction in the number of symptoms, a decrease in severity of any, all, or substantially all of the symptoms, a complete resolution of any, all, or substantially all symptoms, or preventing the occurrence of any, all, or substantially all symptoms associated with one or more of the diseases associated with the use of smoking tobacco as a risk factor, including, but are not limited to, cardiovascular disease, cerebrovascular disease, peripheral vascular disease, aortic aneurysms, emphysema, esophageal cancer, lung cancer, and squamous cell cancers of the head and neck. Tobacco-related diseases also include diseases that are associated with the use of chewing tobacco, such as squamous cell cancers of the mouth.

[0721] In one aspect, the present invention relates to methods of removing senescence associated factors and / or extracellular vesicles from the circulation of a subject in need thereof, thereby de-repressing immune suppression present and / or stimulating regenerative activities in said subjects. Accordingly, the present invention teaches the use of various extracorporeal devices and methods of producing extracorporeal devices for use in clearing senescence associated factors and / or extracellular vesicle content in subjects in need thereof. Said senescence associated factors and / or extracellular vesicles may be elaborated by the senescent tissue itself, or may be generated by surrounding cells under the influence of senescent cells soluble or contact dependent interactions. Said senescence associated factors and / or microvesicles may be directly suppressing the host immune system through induction of T cell apoptosis, proliferation inhibition, incapacitation, anergy, deviation in cytokine production capability or cleavage of the T cell receptor zeta chain, or alternatively saidsenescent associated factors and / or extracellular vesicles may be indirectly suppressing the immune system through modification of function of other immunological cells such as dendritic cells, NK cells, NKT cells and B cells. Said senescence associated factors and / or extracellular vesicles may suppress host regenerative activity through production of fibrosis, inflammatory compounds, and oxidative stress.

[0722] One of the objects of the present invention is to provide an effective and relatively benign treatment for aging associated organ disfunction.

[0723] Another object is to provide an adjuvant, and / or neoadjuvant therapy to be used in conjunction with currently used regenerative medicine treatments. In some cases regenerative medicine treatments that require a functional host immune response for efficacy.

[0724] Another object is to provide an adjuvant, and / or neoadjuvant therapy to be used in conjunction with currently used treatments that stimulate the immune response of a subject in need thereof in an antigen-specific manner.

[0725] Another object is to provide an adjuvant, and / or neoadjuvant therapy to be used in conjunction with currently used treatments that stimulate the immune response of a subject in need thereof in an antigen-nonspecific manner.

[0726] Another object is to provide improvements in extracorporeal treatment of senescence through selecting the novel target of senescence associated microvesicles.

[0727] Another object is to provide beads or other types of particles that can form a matrix outside of a hollow fiber filter, said matrix component having a size greater than pores of said hollow fiber filter, and said beads or other types of particles being bound to agents that capture microvesicles.

[0728] Another object is to provide improvements in extracorporeal treatment of senescence through selecting the novel target of senescent cell associated microvesicles containing unique properties that are not found on microvesicles derived from nonsenescent cells.

[0729] Another object is to provide improved specific affinity devices, particularly immunoadsorption devices, and methods useful for removal of cancer associatedmicrovesicles from cancer patients. Specifically, immunoadsorption devices use proteins with affinity to components of the senescence and / or tumor associated microvesicles. Said proteins include antibodies such as antibodies to Fas ligand, MHC I, MHC II, CD44, placental alkaline phosphatase, TSG-101, MHC I-peptide complexes, MHC II-peptide complexes, CD63, CD81, CD9, EphA2, Integrin a5pi, Integrin aVp3, HLA-G, Notchl, Caveolin-1 , or proteins found to be present on the exterior of microvesicles contributing to senescence and induction of senescence. Contemplated within the invention are proteins that act as ligands for the microvesicular proteins, said proteins may be currently in existence, or may be generated by in silico means based on known qualities of microvesicle-specific proteins.

[0730] In accordance with one particular aspect of the present invention, methods and devices for treating cancer are provided that are based on the utilization of specific affinity adsorption of senescent associated factors and / or extracellular vesicles that are associated with the senescent state. The affinity adsorbents utilized in accordance with the present invention are both immunoadsorbents and non-immune-based specific affinity chemical adsorbents. More specifically, adsorption can be accomplished based on specific properties of the senescent associated microvesicles, one said property is preferential affinity to lectins and other sugar-binding compounds.

[0731] In one particular embodiment, the invention provides a device for extracorporeal treatment of blood or a blood fraction such as plasma. This device has a sorbent circulation circuit, which adheres to and retains senescence associated factors and / ormicrovesicles, and a blood circulation circuit through which blood cells flow unimpeded. The device may be constructed in several variations that would be clear to one skilled in the art. Specifically, the device may be constructed as a closed system in a manner that no accumulating reservoir is needed and the sorbent circulation system accumulates the microvesicles, while non-microvesicle matter is allowed to flow back into the blood circulation system and subsequently returned to the patient. Alternatively, the device may use an accumulator reservoir that is attached to the sorbent circulation circuit and connected in such a manner so that waste fluid is discarded, but volume replenishing fluid is inserted back into the blood circulation system so the substantially microvesicle purified blood that is reintroduced to said patient resembles a hematocrit of significant homology to the blood that was extracted from said patient.

[0732] For the purposes of advancing and clarifying the principles of the invention disclosed herein, reference will be made to certain embodiments and specific language will be used to describe said embodiments. It will nevertheless be understood and made clear that no limitation of the scope of the invention is thereby intended. The alterations, further modifications and applications of the principles of the invention as described herein serve only as specific embodiment, however one skilled in the art to which the invention relates will understand that the following are indeed only specific embodiments for illustrative purposes, and will derive similar types of applications upon reading and understanding this disclosure.

[0733] In accordance with one aspect of the present invention, there are provided methods of removing senescent associated factors and / or extracellular particles from a subject in need thereof, said methods comprising: a) establishing an extracorporeal circulation system which comprises contacting the whole blood or components thereof with a single or plurality of agents capable of binding senescence associated particles and / or extracelluar vesicles found within said blood or components thereoaf; and b) returning said blood or components thereof into the original blood, said blood or blood components containing substantially less senescence associated and / or immune suppressive particles in comparison to the blood or blood components originally residing in the blood.

[0734] Invention methods are useful, for example, for de-repressing immune response, which includes restoration of one or more of the following: T cell, natural killer (NK) cell, natural killer T (NKT) cell, gamma-delta T cell, and B cell function. Presently preferred applications of invention methods include restoration of one or more of the following: T cell, natural killer (NK) cell, natural killer T (NKT) cell, gamma-delta T cell, and B cell function includes prevention of apoptosis; it is especially preferred that restoration of one or more of the following: T cell, natural killer (NK) cell, natural killer T (NKT) cell, gamma-delta T cell, and B cell function includes restoration and / or endowment of activity capable of inhibiting cancer progression.

[0735] Inhibiting cancer progression as contemplated herein is accomplished in a variety of ways, e.g., by one or more of the following: direct cytolysis of tumor cells, direct induction of tumor cell apoptosis, induction of tumor cell cytolysis through stimulation of intrinsic host antitumor responses, induction of tumor cell apoptosisthrough stimulation of intrinsic host antitumor responses, inhibition of tumor cell metastasis, inhibition of tumor cell proliferation, and induction of senescence in the tumor cell.

[0736] Exemplary tumor cells contemplated for treatment herein are selected from the group of cancers consisting of: soft tissue sarcomas, kidney, liver, intestinal, rectal, leukemias, lymphomas, and cancers of the brain, esophagus, uterine cervix, bone, lung, endometrium, bladder, breast, larynx, colon / rectum, stomach, ovary, pancreas, adrenal gland and prostate.

[0737] Agents capable of binding senescent associated proteins / factors and / or extracellular vesicales including microvesicles contemplated for use herein are selected from the group consisting of one or more of the following: a) a singular or plurality of antibody species; b) a singular or plurality of proteins (e.g., lectins); c) a singular or plurality of aptamers, d) a surface that selectively restricts microvesicles from passage, and e) a surface with selective adhesion to microvesicles.

[0738] Antibodies contemplated for use herein have a specificity for proteins selected from the group consisting of one or more of the following: interleukin- 1 beta, interleukin-6 Fas ligand, MHC I, MHC II, CD44, placental alkaline phosphatase, TSG- 101, MHC I-peptide complexes, MHC Il-peptide complexes, CD63, CD81, CD9, EphA2, Integrin a.5|31 , Integrin aV[33, HLA-G, Notchl, Caveolin-1 and proteins found to be present in patients having senescent cells.

[0739] Binding proteins contemplated for use herein are selected from the group comprising consisting of one or more of the following: Fas, T cell Receptor, protein extracts isolated from T cells, protein extracts isolated from dendritic cells, and proteins found to possess affinity for binding proteins found on microvesicles associated with senescent.

[0740] Surfaces contemplated for use herein that selectively restrict passage of said microvesicles typically have pore sizes in the range of about 20-400 nanometers in size, with surfaces having pore sized in the range of about 40-300 nanometers in size being preferred, with surfaces having a pore size in the range of about 50-280 nanometers in size being especially preferred. Surfaces with selective adhesion to soluble senescence associated factors and extracellular vesicles contemplated for use herein can be coatedwith a single compound, or a plurality of compounds that bind particles that are enriched in sphingomyelin and with a lower level of phosphatidylcholine as found in the cellular membranes of non-malignant cells.

[0741] In accordance with another aspect of the present invention, agents capable of binding microvesicles are immobilized on a porous hollow fiber membrane. For example, agents capable of binding microvesicles are immobilized on the porous exterior of the hollow fiber membrane.

[0742] In accordance with another aspect of the present invention, existing methods and devices of extracorporeal treatment of blood can be integrated (in whole or in part) with the above-described methods to augment ex vivo clearance of microvesicles in a physiologically applicable manner. For example, existing methods for extracorporeal treatment of blood can be selected from one or more of the following: a) hemofiltration; b) hemodialysis; and c) hemodiafiltration. A presently preferred existing method for extracorporeal treatment of blood comprises apheresis followed by filtration.

[0743] In accordance with another embodiment of the present invention, there are provided medical devices useful for the removal of senescence associated factors and extracellular vesicles, said device comprising:

[0744] a) an intake conduit through which blood of a patient in need of treatment enters;

[0745] b) a single or plurality of matrices capable of adhering to extracellular vesicles causative of senescent associated immune suppression; and

[0746] c) a system for reintroduction of said blood into the patient in need thereof, whereby said blood is reintroduced under physiologically acceptable conditions.

[0747] In one aspect of the above-described medical device, the matrices surround a plurality of hollow fiber filters. Preferably, the hollow fiber filters have a diameter of sufficient size to allow passage of blood cells through the lumen, and diffusion of particles between 80-300 nanometers in size.

[0748] In another aspect for the above-described medical device, a microvesicle binding agent is chemically reacted with a high-molecular weight substrate and placed onthe exterior of said hollow fibers so as to bind non-blood cell liquids permeating through the pores of said hollow fibers. Exemplary microvesicle binding agents include one or more of the following: a) a singular or plurality of antibody species; b) a singular or plurality of proteins (e.g., lectins); c) a singular or plurality of aptamers, d) a surface that selectively restricts microvesicles from passage, and e) a surface with selective adhesion to microvesicles.

[0749] Exemplary antibodies contemplated for use herein have a specificity for proteins selected from the group consisting of one or more of the following: Interleukin- 1 beta Fas ligand, MHC I, MHC II, CD44, placental alkaline phosphatase, TSG-101, MHC I-peptide complexes, MHC Il-peptide complexes, CD63, CD81, CD9, EphA2, Integrin a5pi, Integrin aV[33, HLA-G, Notchl, Caveolin-1 and proteins found to be present on the exterior of extracellular contributing to immune suppression found in a patient possessing senescent cells. Presently preferred antibodies are specific to Fas ligand, MHC I, and HLA-G.

[0750] Exemplary proteins contemplated for use in the invention device are selected from the group consisting of one or more of the following: Fas, T cell Receptor, protein extracts isolated from T cells, protein extracts isolated from dendritic cells, and proteins found to possess affinity for binding proteins found on extracellular vesicles associated with senescence.

[0751] In accordance with another embodiment of the present invention, there are provided methods of potentiating the immunologically mediated regenerative response elicited by vaccination to senescence associated antigens, said methods comprising:

[0752] a) immunizing a subject in need thereof using a single or combination of senescence associated antigens;

[0753] b) removing senescence associated factors and / or from the sera of said subject by extracorporeal means; and

[0754] c) adjusting the amount of removal of immune suppressive / degeneration promoting extracellular vesicles based on immune stimulation / regeneration desired. In accordance with yet another embodiment of the present invention there are provided methods of enhancing the regeneration stimulation activity of a subject in need thereofthrough the removal of senescence associated extracellular vesicular particles found in systemic circulation of said subject, said methods comprising:

[0755] a) establishing an extracorporeal circulation system which comprises contacting the whole blood or components thereof with a single or plurality of agents capable of binding microvesicles found within said blood or components thereof; and b) returning said blood or components thereof into the subject, said blood or blood components containing substantially less immune suppressive / degenerative factors / particles in comparison to the blood or blood components originally residing in said subject.

[0756] Enhancing immune response as contemplated herein includes one or more of the following: upregulation of T cell, natural killer (NK) cell, natural killer T (NKT) cell, gamma-delta T cell, and B cell function. In a presently preferred embodiment, upregulation of one or more of T cell, natural killer (NK) cell, natural killer T (NKT) cell, gamma-delta T cell, and B cell function includes prevention of apoptosis. In yet another preferred embodiment, upregulation of one or more of T cell, natural killer (NK) cell, natural killer T (NKT) cell, gamma-delta T cell, and B cell function includes enhancing and / or endowment of activity capable of inhibiting cancer progression.

[0757] In some embodiments the invention provide suppression of cancer by removal of cancer induced senescence associated factors and / or extracellular vesicles.Inhibiting cancer progression contemplated herein is accomplished in a variety of ways, e.g., by direct cytolysis of tumor cells, direct induction of tumor cell apoptosis, induction of tumor cell cytolysis through stimulation of intrinsic host antitumor responses, induction of tumor cell apoptosis through stimulation of intrinsic host antitumor responses, inhibition of tumor cell metastasis, inhibition of tumor cell proliferation, and induction of senescence in the tumor cell. Tumor cells contemplated for treatment in accordance with the present invention are selected from the group of cancers consisting of soft tissue sarcomas, kidney, liver, intestinal, rectal, leukemias, lymphomas, and cancers of the brain, esophagus, uterine cervix, bone, lung, endometrium, bladder, breast, larynx, colon / rectum, stomach, ovary, pancreas, adrenal gland and prostate. Agents capable of binding microvesicles contemplated for use herein are selected from the group consisting of one or more of the following: a) a singular or plurality of antibody species; b) a singular or plurality of proteins (e.g., lectins); c) a singular or plurality of aptamers, d) asurface that selectively restricts microvesicles from passage, and e) a surface with selective adhesion to microvesicles. Antibodies having specificity for proteins contemplated for use herein are selected from the group consisting of one or more of the following: Fas ligand, MHC I, MHC II, CD44, placental alkaline phosphatase, TSG-101, MHC I-peptide complexes, CD63, CD81, CD9, EphA2, Integrin a5pi, Integrin aVp3, HLA-G, Notch 1, Caveolin-1 MHC II-peptide complexes, and proteins found to be present on the exterior of microvesicles contributing to senescnece

[0758] Proteins contemplated for use herein are selected from the group consisting of one or more of the following: Fas, T cell Receptor, protein extracts isolated from T cells, protein extracts isolated from dendritic cells, and proteins found to possess affinity for binding proteins found on microvesicles associated with immune suppression. Surfaces that selectively restrict passage of said microvesicles contemplated for use herein typically have a pore size in the range of about 20-400 nanometers in size, with pores sizes in the range of about 40-300 nanometers in size being preferred, and pore sizes in the range of about 50-280 nanometers in size being especially preferred.

[0759] Surfaces with selective adhesion to microvesicles contemplated for use herein are coated with a variety of agents, e.g., a single compound, or plurality of compounds that bind particles that are enriched in sphingomyelin and with a lower level of phosphatidylcholine as found in the cellular membranes of non-malignant cells. In one aspect, the above-described agents capable of binding microvesicles are immobilized on a porous hollow fiber membrane, e.g., on the porous exterior of the hollow fiber membrane.

[0760] In another aspect of the invention, existing methods and devices of extracorporeal treatment of blood are integrated (in whole or in part) for augmenting ex vivo clearance of microvesicles in a physiologically applicable manner. Exemplary existing methods for extracorporeal treatment of blood are selected from one or more of the following: a) hemofiltration; b) hemodialysis; and c) hemodiafiltration. A presently preferred existing method for extracorporeal treatment of blood comprises apheresis followed by filtration.

[0761] In accordance with yet another embodiment of the present invention, there are provided methods of enhancing the immune response of a subject in need thereof through the removal of microvesicular particles found in systemic circulation of saidsubject, said methods comprising: a) establishing an extracorporeal circulation system which comprises contacting the whole blood or components thereof with a single or plurality of agents capable of binding microvesicles found within said blood or components thereof, said agents being in turn bound to a plurality of objects; b) performing a filtration step such that said objects of a defined size are captured within said extracorporeal circulation system; and c) returning said blood or components thereof into the subject, said blood or blood components containing substantially less immune suppressive particles in comparison to the blood or blood components originally residing in said subject.

[0762] Enhancing immune response contemplated herein includes one or more of the following: upregulation of T cell, natural killer (NK) cell, natural killer T (NKT) cell, gamma-delta T cell, and B cell function. It is presently preferred that upregulation of one or more of T cell, natural killer (NK) cell, natural killer T (NKT) cell gamma-delta T cell, and B cell function includes prevention of apoptosis. It is also presently preferred that upregulation of one or more of T cell, natural killer (NK) cell, natural killer T (NKT) cell gamma-delta T cell, and B cell function includes enhancing and / or endowment of activity capable of inhibiting cancer progression.

[0763] Inhibiting cancer progression contemplated herein is accomplished by one or more of the following: direct cytolysis of tumor cells, direct induction of tumor cell apoptosis, induction of tumor cell cytolysis through stimulation of intrinsic host antitumor responses, induction of tumor cell apoptosis through stimulation of intrinsic host antitumor responses, inhibition of tumor cell metastasis, inhibition of tumor cell proliferation, and induction of senescence in the tumor cells.

[0764] Tumor cells contemplated for treatment in accordance with the present invention are selected from the group of cancers consisting of: soft tissue sarcomas, kidney, liver, intestinal, rectal, leukemias, lymphomas, and cancers of the brain, esophagus, uterine cervix, bone, lung, endometrium, bladder, breast, larynx, colon / rectum, stomach, ovary, pancreas, adrenal gland and prostate. Agents capable of binding microvesicles contemplated for use herein are selected from the group consisting of one or more of the following: a) a singular or plurality of antibody species; b) a singular or plurality of proteins (e.g., lectins); c) a singular or plurality of aptamers, d) asurface that selectively restricts microvesicles from passage, and e) a surface with selective adhesion to microvesicles.

[0765] The plurality of objects contemplated for use herein comprise beads manufactured to a specific size or range of sizes in a manner so that said agents capable of binding microvesicles may be conjugated to said plurality of objects. Preferably such beads have a defined size range to restrict their movement out of said extracorporeal circulation system, e.g., the beads are of a size range larger than pores of hollow fibers used in extracorporeal systems so as to restrict their movement out of said extracorporeal systems. In one aspect, such beads possess properties responsive to an electromagnetic field, such that subsequent to said beads contacting said microvesicles, said beads may be removed or sequestered by said electromagnetic field in order to substantially prevent movement of said beads out of said extracorporeal system. Examples of beads contemplated for use herein are MACS™ beads alone or conjugated with compounds in order to allow said beads to form complexes with said agents capable of binding microvesicles, Dynal™ beads alone or conjugated with compounds in order to allow said beads to form complexes with said agents capable of binding microvesicles, and the like.

[0766] Antibodies contemplated for use herein have a specificity for proteins selected from a group consisting of one or more of the following: Fas ligand, MHC I, MHC II, CD44, placental alkaline phosphatase, TSG-101, MHC I-peptide complexes, MHC II-peptide complexes, CD63, CD81, CD9, EphA2, Integrin a5[31, Integrin aV[33, HLA-G, Notch 1, Caveolin-1 and proteins found to be present on the exterior of microvesicles contributing to senesnce. Presently preferred antibodies are specific to Fas ligand, MHC I, and the like. Proteins contemplated for use herein are selected from the group consisting of one or more of the following: Fas, T cell Receptor, protein extracts isolated from T cells, protein extracts isolated from dendritic cells, and proteins found to possess affinity for binding proteins found on microvesicles associated with immune suppression. Surfaces that selectively restrict passage of said microvesicles typically fall in the range of about 20-400 nanometers in size, with microvesicles falling in the range of about 40-300 nanometers in size being presently preferred, and microvesicles in the range of about 50-280 nanometers in size being especially preferred. Exemplary surfaces with selective adhesion to microvesicles are coated with a single compound, or plurality of compounds that bind particles that are enriched in sphingomyelin and with a lower levelof phosphatidylcholine as found in the cellular membranes of non-malignant cells. In accordance with another aspect of the invention, agents capable of binding microvesicles can be immobilized on a porous hollow fiber membrane, e.g., on the porous exterior of the hollow fiber membrane. n accordance with still another aspect of the present invention, existing methods and devices of extracorporeal treatment of blood can be integrated (in whole or in part) with the above-described methods to augment ex vivo clearance of microvesicles in a physiologically applicable manner. Exemplary methods contemplated for use herein include: a) hemofiltration; b) hemodialysis; and c) hemodiafiltration, with a preferred method including apheresis followed by filtration.

[0767] In accordance with various aspects of the present invention, extracorporeal removal of microvesicles can be performed through selective adhesion of said microvesicles to matrices or substrates that are conjugated to agents possessing higher affinity to microvesicles with a high sugar content, in comparison to microvesicles of a lower sugar content. In accordance with yet another embodiment of the present invention, there are provided methods of extracorporeally removing microvesicles from a subject in need thereof, said method comprising passing said subject's whole blood, or separated blood components, through a system capable of selectively binding and retaining microvesicles based on one or more of size, charge, affinity towards lectins, or affinity towards molecules that are known to be present on said microvesicles. In accordance with a further embodiment of the present invention, there are provided methods of extracorporeally removing microvesicles from a subject in need thereof, said methods comprising passing said subject's whole blood, or separated blood components, through a system capable of non-selectively binding and retaining microvesicles based on one or more of size, charge, affinity towards lectins, or affinity towards molecules that are known to be present on said microvesicles. In accordance with a still further embodiment of the present invention, there are provided methods of extracorporeally removing microvesicles from a subject in need thereof, said methods comprising passing said subject's whole blood, or separated blood components, through a system capable of selectively binding and retaining microvesicles based on similarities between properties of microvesicles and membranes of cancer cells. In accordance with yet another embodiment of the present invention, there are provided methods of extracorporeally removing microvesicles from a subject in need thereof, said methods comprising passing said subject's whole blood, or separated blood components, through a system capable ofnon- selectively binding and retaining microvesicles based on similarities between properties of microvesicles and membranes of senescent cells and / or cancer induced senescent cells.

[0768] In some embodiments of the invention, removement of systemic senescence associated factors and / or extracellular vesicles is used together with agents that suppress senescence. One or more of the agents of the invention (e.g., peptides, polypeptides, proteins, small molecules, antibodies, or antibody fragments that target senescent cells) can also be used to prepare compositions for therapeutic administration (e.g., to treat, stabilize, inhibit the development or progression of, or prevent age-related diseases, tobacco-related diseases, cancer, neurodegenerative diseases, and other diseases and disorders related to or caused by cellular senescence) by coupling of the agents of the invention to, e.g., therapeutic and / or cytotoxic agents. Senescent cells have been observed in skin to make up only a fraction of the stromal cell compartment even in old donors (Dimri et al., Proc. Nat. Acad. Sci. USA 92:9363-9367, 1995) and up to 16% of stromal fibroblasts in patients with emphysema (Bergner, Respiratory Res. 7:32, 2006), but senescent cells can exert clinically significant paracrine effects even when they compose only 10% of the content of stromal cells (Krtolica et al., Proc. Nat. Acad. Sci. USA 98: 12072-12077, 2001). Thus, eliminating senescent cells while leaving intact the majority of the cellular compartment from which the senescent cells originate can ablate the harmful paracrine effects of senescent cells. The secretion of elevated levels of collagenase and elastase and depressed levels of collagenase inhibitors by senescent stromal cells implicates them in diseases or conditions that feature breakdown or decrease in structural integrity of the extra cellular matrix, including emphysema, aortic aneurysms, vascular disease, osteoarthritis, and skin wrinkling. Elimination of some, all, or substantially all senescent stromal cells in a patient (e.g., in a tissue or organ of a patient) can attenuate the progression, decrease the symptoms, or prevent the occurrence of these diseases and conditions. Senescent stromal cells have the ability to stimulate tumorigenesis and, by weakening the extracellular matrix, facilitate metastasis. Thus, the elimination of some, all, or substantially all of the senescent stromal cells in a patient (e.g., in a tissue or organ of a patient) can decrease the risk of tumorigenesis and metastasis (e.g., by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99% or more).

[0769] The senescent cell targeting agents of the invention (e.g., peptides, polypeptides, proteins, small molecules, antibodies, or antibody fragments that target senescent cells) can be prepared by amino acid coupling using solid phase peptide synthesis (SPPS). As is known in the art, the amino acids to be used as substrates to form the agents of the invention are Fmoc-protected prior to incorporation into a peptide chain (see, e.g., Chanand White, FMOC Solid Phase Peptide Synthesis, A Practical Approach, Oxford University Press, New York, 2003); incorporated herein by reference in its entirety). The standard coupling techniques used to couple the amino acids in order to form the agents of the invention are known in the art (see, e.g., Chan and White, supra). For example, a polyamide-Rink resin can be prepared by loading a polyamide resin with Fmoc-Rink using chemical protocols well known in the art (see, e.g., Chan and White, supra). The first amino acid in the peptide sequence is coupled to the resin after removing Fmoc from the N-terminal amine of the resin using piperidine. Once the coupling is complete, the resin is washed and Fmoc is removed from the coupled amino acid using piperidine. The resin is washed again, and the next amino acid in the sequence is coupled to the previously coupled amino acid. This process is repeated using the necessary amino acids until the desired peptide is formed. Following the coupling of the final amino acid, the Fmoc group is removed using piperidine. The terminal amine can be left as a free amine, or it can be acetylated. The peptide can be cleaved from the resin using trifluoroacetic acid (TFA), triisopropylsilane, and water according to techniques known in the art (see, e.g., Chan and White, supra). The cleaved peptide is then separated from the residue by filtration. The TFA is typically evaporated to dryness followed by precipitation of the peptide with diethyl ether. Typically, the final peptide product is purified using HPLC. Mass spectrometry is used to verify that the desired peptide is obtained. The agents of the invention (e.g., peptides, polypeptides, proteins, small molecules, antibodies, or antibody fragments that target senescent cells) can be readily prepared by automated solid phase peptide synthesis using any one of a number of well-known, commercially available automated synthesizers, such as the Applied Biosystems ABI 433A peptide synthesizer.

[0770] The agents of the invention (e.g., peptides, polypeptides, proteins, small molecules, antibodies, or antibody fragments that target senescent cells) can be labeled for fluorescence detection by labeling the agent with a fluorophore, such as rhodamine or fluorescein, using techniques well known in the art (see, e.g., Lohse et al., Bioconj.Chem. 8:503-509, 1997). The senescent cell targeting agents of the invention (e.g., peptides, polypeptides, proteins, small molecules, antibodies, or antibody fragments that target senescent cells) can also be labeled with a radioactive metal or a relaxivity metal by coupling the agent to a metal chelating agent that chelates a radioactive metal or relaxivity metal. Examples of chelating agents include, but are not limited to, ininocarboxylic and polyaminopolycarboxylic reactive groups, di ethylenetriaminepentaacetic acid (DTP A), and 1,4,7, 10-tetraazacyclododecane- 1,4,7, 10-tetraacetic acid (DOTA). The chelating agent can be coupled via its amino acid side chain directly to the agents of the invention. Alternatively, an intervening amino acid sequence can be coupled using SPPS to both the agents of the invention and the chelating agent.

[0771] When carrying out the above-described methods, the similarities between cancer associated microvesicles and membranes of senescent and / or cancer induced senescent cells include ability to bind a lectin or plurality of lectins. Reference to lectins herein includes GNA, NPA, Conconavalin A and cyanovirin, with a presently preferred lectin being Conconavalin A. One embodiment of the present invention relates to methods that can be used for extracorporeal treatment of blood or a blood fraction for the removal of microvesicles associated with immune suppression in a cancer patient. Blood is run through an extracorporeal circulation circuit that uses a hollow fiber cartridge with the membranes of said hollow fibers having sufficient permeability for the microvesicles found in the blood to be removed through the membrane of the hollow fibers and into an area outside of the fibers containing a substrate that is bound to a single or plurality of agents capable of adhering to said microvesicles in a manner such that said microvesicles are attached to said agent and do not substantially re-enter the hollow fibers. Within the knowledge of one skilled in the art are available numerous types of hollow fiber systems. Selection of said hollow fiber system is dependent on the desired blood volume and rate of passage of said blood volume through the hollow fiber system. Specifically, hollow fiber cartridges may be used having lengths of 250 mm and containing 535 hollow fibers supplied by Amicon, and having the fiber dimensions: I.D. 180 micron and O.D. 360 micron, and the total contact surface area in the cartridge is 750 cm2. Alternatively, the “Plasmaflux P2” hollow fiber filter cartridge (sold by Fresenius) or Plasmart PS60 cartridges (sold by Medical srl) may be used. These and other hollow fiber systems are described by Ambrus and Horvath in U.S. Pat. No. 4,714,556 and incorporated herein byreference in its entirety. Hollow fiber cartridges such as described by Tullis in United States Patent Application 20040175291 (incorporated by reference herein in its entirety) may also be used. Furthermore, said hollow fiber cartridges and affinity cartridges in general are thought in U.S. Pat. Nos. 4,714,556, 4,787,974 and 6,528,057, which are incorporated herein by reference in their entirety.

[0772] Regardless of hollow fiber system used, the concept needed for application of the present invention, is that said hollow fiber filters are required to allow passage of blood cells through the interior of said hollow fiber, and allow diffusion of microvesicles to the exterior. In order to allow such diffusion, the pores on the membrane of the hollow fiber need to be of a diameter sufficient to allow particles ranging from the size of 20 nanometers to 500 nanometers in diameter. More specifically, the pores on the membrane of the hollow fiber need to be of a diameter sufficient to allow particles ranging from the size of 50 nanometers to 300 nanometers in diameter. Even more specifically, the pores on the membrane of the hollow fiber need to be of a diameter sufficient to allow particles ranging from the size of 80 nanometers to 200 nanometers in diameter. During experimentation with different hollow fibers, one skilled in the art would find it useful to utilize particles of similar size ranges as the microvesicles in order to calibrate and quantitate the ability of various pore sizes of hollow filters. One method of performing this is through the utilization of commercially available MACS™ Beads (Milteny Biotech), which have a size of 60 nanometers. Fluorescent, spherical latex beads ranging in size from 25 to 1000 nm are also available for this purpose (e.g., from Duke Scientific (Palo Alto, Calif.)). The substrate or matrix to be used in practicing the present invention needs to allow sufficient permeation of flow so that non-cellular blood components that enter the space exterior to the hollow fiber are distributed throughout the substrate or matrix material, so that substantial contact is made between the microvesicles permeating the hollow fiber filter and the microvesicle-binding agent that is attached to the substrate or matrix. Suitable substrates or matrices are known to one skilled in the art. Said substrates or matrices include silica gel, dextran, agarose, nylon polymers, polymers of acrylic acid, co-polymers of ethylene and maleic acid anhydride, aminopropylsilica, aminocelite, glass beads, silicate containing diatomaceous earth or other substrates or matrices known in the art. Examples of such are described in the following patents, each of which are incorporated by reference herein in their entirety: Lentz U.S. Pat. No. 4,708,713, Motomura U.S. Pat. No. 5,667,684, Takashima et al U.S. Pat. No. 5,041,079,and Porath and Janson U.S. Pat. No. 3,925,152. The agents that are attached to said substrate are chosen based on known affinity to cancer associated microvesicles. Said agents may be capable of non-specifically binding to said microvesicles, in that binding occurs both from non-tumor associated microvesicles, and from tumor associated microvesicles, or conversely, said agents may display a certain degree of selectivity for exosomes derived from tumors. In one embodiment said agents non-specifically bind all microvesicles due to common expression of molecules such as MHC I on microvesicles that are associated with conditions of neoplasia, and microvesicles that are not. Specifically, an agent that would bind both types of microvesicles would be an antibody specific to the non-polymorphic regions of MHC I. Therefore, in the embodiment of the invention in which non-selective removal of microvesicles is sought, anti-MHC I antibodies would be bound to said substrate chosen, and the combination would be placed to reside outside of the hollow fiber filters in order to allow binding of said microvesicles to the substrate, however blood cells and other components of the blood would not be removed during the passage of blood through the encased system containing said hollow fiber filters, exterior substrate, and microvesicle binding agent.

[0773] In order to achieve non-specific removal of microvesicles, another embodiment of the invention is the use of hollow fiber filters of sufficient size of the pores on the side of the hollow fiber filter for microvesicles to exit, while not allowing blood cells to exit, and passing a continuous solution over said hollow fiber filters in order to clear said microvesicles leaking through the sides of the hollow fiber filters. In such a situation it would be critical to re-introduce the other blood components that escaped the hollow fiber filter, such as albumin, back into the microvesicle purified blood, before returning of the blood to the subject.

[0774] Alternatively, the hollow-fiber cartridge may be sealed as decribed in Ambrus. In such a system, both diffusion and convection cause blood fluids (exclusive of blood cells) to pass through the pores in the hollow fibers and into contact with the capture molecules bound to the solid phase matrix. The fluids (e.g. plasma) pass back into the circulation at the distal end of the cartridge through a process known as Starling flow. In this system, there is no significant loss of blood fluids and therefore no need for blood component replacement. In the situations where a substantially specific removal of microvesicles associated with tumors is desired, the said agent bound to said substrateoutside of said hollow fiber filters possesses affinity to molecules specifically found on said microvesicles associated with tumors. Said agent may be an antibody to the molecule Fas Ligand, may be a recombinant Fas protein, or may be directed to MHC I, MHC II, CD44, CD63, CD81, CD9, EphA2, Integrin a5 1, Integrin aVp3, HLA-G, Notchl, Caveolin-1 placental alkaline phosphatase, TSG-101, MHC I-peptide complexes, MHC I- peptide complexes, and proteins found to be present on the exterior of microvesicles contributing to senescence.

[0775] In a situation where microvesicles associated with tumors are meant to be withdrawn with a certain degree of selectivity from the systemic circulation of a subject in need thereof, said agent binding the matrix or substrate may be a lectin. Specific methodologies for use of lectins in removal of viruses are described by Tullis in United States Patent Application 20040175291 (incorporated by reference herein in its entirety) and these methodologies may also be used in part or in whole for practicing the present invention. In various embodiments of the invention, it is important that said systems include means for maintaining the blood at conditions similar to that found in the host, so that upon returning said blood to the host, no adverse reactions occur. In other words, it is within the scope of the invention to use technologies that are known to one skilled in the art to maintain blood at physiological ion concentrations, osmolality, pH, hematocrit, temperature, and flow in order to avoid harm being caused to the subject subsequent to reinfusion of blood treated as disclosed herein. Said technologies are well known to one skilled in the art.

[0776] In another embodiment of the invention a system for extracorporeal clearance of microvesicles; either selectively removing tumor associated microvesicles, or non- selectively microvesicles that are found in healthy subjects as well as tumor bearing subjects. The invention comprises several interacting components whose primary purpose is the formation of a functional circuit capable of depleting microvesicles in order to derepress, or in some cases augment the immune response of a cancer patient. More specifically, a means for separating blood from a subject in need thereof (e.g., a cancer patient) into plasma and cellular elements is used. Appropriate means for such separation are available commercially, and well-known to the skilled artisan. They include, for example, the Exorim System, the Fresenius Hemocare Apheresis system, and the Gambo Prisma System. Plasma purified through said separation means is then run over an arrayof filtration means, said filtration means possessing a higher affinity towards tumor associated microvesicles in comparison to other molecules. Said filtration means includes, in some embodiments, microvesicle binding agents immobilized to a substrate. Said microvesicle binding agents include but are not limited to antibodies, proteins, or compounds with selective affinity towards microvesicles associated with the cancer or not associated. Examples of such agents include antibodies to Fas ligand, MHC I, MHC II, CD44, placental alkaline phosphatase, TSG-101, MHC I-peptide complexes, CD63, CD81, CD9, EphA2, Integrin a5pi, Integrin aVp3, HLA-G, Notchl, Caveolin-1 MHC II-peptide complexes, or proteins found to be present on the exterior of microvesicles contributing to senescence as well as lectins such as conconavalin A, phytohemagluttanin, GNA, NPA, and cyanovirin. Said substrate is selected from known substrates previously used in the are, these include, for example SEPHAROSE™ made by Amersham- Biosciences, Upsala, Sweden, as well as acrylamide and agarose particles or beads. The substrates used should have the properties of being able to tightly bind the microvesicle binding agent, the ability to be produced in a sterile means, and be compatible with standard dialysis / extracorporeal tubings.

[0777] In other embodiments, the agent capable of binding the tumor associated or non-tumor associated microvesicles is immobilized to a filter membrane or capillary dialysis tubing, where the plasma passes adjacent to, or through, the membranes to which said agent capable of binding the tumor associated or non-tumor associated microvesicles are bound. Suitable filters include those mentioned previously with respect to separation of blood components. These may be the same filters, having immobilized agents capable of binding microvesicles (either tumor associated or non-associated, or may be arranged in sequence, so that the first filter divides the blood components and the secondary, tertiary and additional filter removes one or more of the components of said cancer associated microvesicles. Conjugation of the agent capable of binding the tumor associated or non-associated microvesicle to said substrate may be accomplished by numerous means known in the art. Said means include avidin-streptavidin, cynanogen bromide coupling, the use of a linker such as a polyethylene glycol linker. A means of returning the blood together with plasma substantially cleared of tumor associated microvesicles back to said subject is also provided in the invention. Preferred means are chosen by one of skill in the art based on the desired application, extent of microvesicle removal desired, patient condition, extracorporeal method chosen, and microvesicle-binding agent chosen. In one embodiment of the invention, extracorporeal removal of microvesicles is performed in a cancer patient in order to accelerate the rate of tumorspecific T cell proliferation and activation. It is known in the art that tumors contain antigens that are specific to the tumor (e.g. the bcr-abl product p210 in CML), expressed on other tissues but overexpressed on cancer cells (e.g. tyrosinase), or expressed embryonically and re-expressed in the cancer (e.g. telomerase). Vaccination to such antigens has been demonstrated to induce immune response, and in some cases generation of cytotoxic T lymphocytes (CTL). Unfortunately, despite much effort in development of cancer vaccines, clinical translation has been slow, with most cancer vaccines not demonstrating efficacy in the double-blind setting. In order to increase efficacy of cancer vaccines, it is important that the cancer patient has an immunological environment in which proper T cell activation may occur. It is known that high numbers of microvesicles are present in the circulation of patients with a wide variety of histologically differing tumors including melanoma (52), ovarian (53), colorectal (54), and breast (55). Importantly, such microvesicles are known to induce suppression of immunity via direct mechanisms such as induction of T cell death via FasL expression (52), through indirect mechanisms such as stimulation of myeloid suppressor cell activity (54). Indeed, numerous mechanisms are known for suppression of T cell immunity by cancer-secreted microvesicles (56-59). Accordingly, in one embodiment a cancer patient is treated with a therapeutic cancer vaccine either prior to, concurrently, or subsequent to undergoing extracorporeal removal of exosomes. Said cancer vaccine may be used for stimulation of immune responses to antigens that are found either exclusively on the tumor, to antigens found on non-malignant tissues but at higher concentration on the tumor, or antigens whose presence is required for tumor functionality. In a specific embodiment of the invention, tumor vaccination is performed to peptides, polypeptides, glycoproteins, peptidomimetics, or combinations thereof. Tumor vaccination may be performed in the context of a cell therapy, such as, for example, administration of dendritic cells that are pulsed with tumor antigens or tumor lysates. Tumor vaccines are commonly known in the art and are described in the following reviews, which are incorporated by reference (60- 64). Examples of tumor antigens that may be used in the practice of the current invention include CDK-4 / ma MUM-1 / 2, MUM-3, Myosin / m, Redox-perox / m, MART-2 / m, Actin / 4 / ma, ELF2-M, CASP-8 / ma, HLA-A2-R17OJ, HSP70-2 / ma, CDKN2A, CDC27a, TPI, LDLR / FUT Fibronectin / m, RT-PTP-K / ma, BAGS, GAGE, MAGE, telomerase, and tyrosinase, and fragments thereof. In one specific embodiment, a patient with ovariancancer is selected for treatment with cancer vaccination. Said patient plasma is assessed for exosomal content based on methods known in the art, as for example described in the following study and incorporated herein by reference (65). In specific method involves the following procedure: ETDA treated plasma is purified from peripheral blood by centrifugation at 500 g for half-hour. Separation of cellular debris is accomplished by a second centrifugation at 7,000 g for an additional half-hour. Exosomes are subsequently collected by centrifugation at 100,000 g for 3 hours, followed by a washing step in PBS under the same conditions. Using this procedure, approximately 0.5-0.6 ug / ml of exosomal protein is detected from healthy volunteers as visualized by the Bradfort Assay (Bio-Rad, Hercules, Calif.) (66). In contrast, the plasma of cancer patients typically contains a higher exosomal yield, ranging between 200-500 ug / ml. This is in agreement with studies describing high concentrations of “membrane vesicles” found systemically circulating in cancer patients (65). For the practice of the invention patients with high exosomal content compared to healthy volunteers are selected. For example, patients with exosomal content above two fold the concentration of exosomes in healthy volunteers may be treated by the invention. In another embodiment, patients with exosomal contented 10-fold higher than exosomal content of healthy volunteers are treated. In another embodiment of the invention, patients with higher exosomal content than healthy volunteers which have spontaneous T cell apoptosis present are selected for treatment. Protocols for assessment of spontaneous T cell apoptosis are known in the art and described, for example by Whiteside's group and incorporated here by reference (67). Assessment of exosome immune suppressive activity may be quantified by culture of exosomes purified from patient plasma with a Fas expressing T cell line such as the Jurkat clone E6.1 (ATCC Manassas, Va.). These cells may be cultured in standard conditions using the method described by Andreola et al and incorporated herein by reference (52) in order to develop a standardized assay. Briefly 106 / ml Jurkat cells are seeded in 24-well plates in 10% FBS RPMI 1640 and co-cultured with escalating concentrations of exosomes from healthy volunteers, as well as cancer patients. Apoptosis of Jurkat cells may be quantified by assessment of Annexin-V staining using flow cytometry.

[0778] Patients displaying elevated numbers of exosomes, and / or apoptotic T cells, and / or possessing exosomes capable of inducing T cell apoptosis are selected for extracorporeal removal of said exosomes. In one preferred embodiment, patent blood ispassaged over an extracorporeal circuit for a time sufficient to substantially reduce exosome burden. Reduction of exosome burden is quantified as described above. Correlation can be made between exosome concentration and spontaneous T cell apoptosis. When reduction of both plasma exosome concentration and spontaneous T cell apoptosis is achieved, said patient may be immunized with a tumor vaccine.Alternatively, patients may have exosome removal performed without immunization with a tumor vaccine so as to allow for endogenous antitumor responses to be derepressed. Alternatively patients may be treated with a non-specific immune stimulator, said immune stimulator may be a small molecule (e.g. muramyl dipeptide, thymosin, 7,8- disubstituted guanosine, imiquimod, detoxified lipopolysaccharide, isatoribine or alphagalactosylceramide), a protein (e.g. IL-2, IL-7, IL-8, IL-12, IL-15, IL-18, IL-21, IL-23, IFN-a, b, g, TRANCE, TAG-7, CEL-1000, bacterial cell wall complexes, or LIGHT), or an immunogeneic nucleic acid (e.g. short interfering RNA targeting the mRNA of immune suppressive proteins, CpG oligonucleotides, Poly IC, unmethylated oligonucleotides, plasmid encoding immune stimulatory molecules, or chromatin-purified DNA). Said non-specific immune stimulants are known in the art and in some cases are already in clinical use. Said non-specific immune stimulants in clinical use include interleukin-2, interferon gamma, interferon alpha, BCG, or low dose cyclophosphamide. In another embodiment extracorporeal removal of exosomes is performed in conjunction with chemotherapy in order to derepress immune suppression caused by exosomes, while at the same time allowing said chemotherapy to perform direct tumor inhibitory functions. Alternatively, extracorporeal removal of exosomes may be utilized to remove increased exosomes caused by tumor cell death during chemotherapy use. Numerous types of chemotherapies are known in the art that may be utilized in the context of the present invention, these include: alkylating agents such as thiotepa and cyclosphosphamide (CYTOXAN.™.); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide and trimethylolomelamine; nitrogen mustards such as chlorambucil, chlomaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomysins, actinomycin, authramycin, azaserine,bleomycins, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5 -fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenishes such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxanes, e.g. paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N.J.) and docetaxel (TAXOTERE®, Rhone- Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylomithine (DMFO); retinoic acid; esperamicins; and capecitabine. In one embodiment, frequency and length of extracorporeal treatment is performed based on the amount of time (or blood volume) needed for reduction of exosome concentration to a level significant to correlate with reduction in spontaneous T cell apoptosis. In one embodiment a reduction of spontaneous T cell apoptosis by approximately 20% in comparison to pre- extracorporeal treatment values is judged as sufficient. In another embodiment a reduction of spontaneous T cell apoptosis by approximately 50% in comparison to pre- extracorporeal treatment values is judged as sufficient. In another embodiment areduction of spontaneous T cell apoptosis by approximately 90% in comparison to pre- extracorporeal treatment values is judged as sufficient.

[0779] Although assessment of spontaneous T cell apoptosis is used in some embodiments forjudging the frequency, and / or time, and / or blood volume needed for extracorporeal treatment, other means of measuring immune responses may be used. For example, restoration of cytokine production, T cell proliferation, or TCR-zeta chain expression are all known in the art and described in the references incorporated herein.

[0780] In one embodiment, removal of senescence associated factors is used together with approaches that stimulate neurogenesis for example electroconvulsive therapy [1, 2], transcranial magnetic stimulation [3-30], low intensity transcranial ultrasound stimulation [31-33], transcranial direct current stimulation

[0034] ,

[0781] In another embodiment removal of senescence associated factors is used together with approaches that suppress inflammation, including approaches that reduce Macrophage 2

[0035] , astrocyte 2, or neutrophil 2 activity.

[0782] In some embodiments removal of senescence associated factors is combined with suppressors of interleukin- 17 for treatment of inflammatory diseases

[0036] , It is known that patients with liver failure have elevated systemic levels of IL- 17 as well as IL-17 made by T cells [37, 38], IL-17 levels correlate with severity of liver failure [39- 43], Levels of Thl7 cells which produce IL-17 correlate with liver failure [44-47], L-17 induces liver failure in animal models [48-51] and patients

[0052] , Suppressing IL-17 reduces liver failure in animal models [53-59],

[0783] In one embodiment the invention describes removal of the senescence associated protein IL- 17 and various upstream inducer proteins. In the case of multiorgan failure, production of IL- 17 during ischemia reperfusion injury stimulates organ failure

[0060] , additionally, association between IL- 17 and organ failure [61, 62], In the situation of heart failure, it is published that blocking IL- 17 reduces autoimmune heart failure [63, 64], IL-17 induces inflammation and pathology in models of heart failure [65-67], IL-17 associated with heart failure in people [68-76], with correlation between IL- 17 and severity of cardiac dysfunction. Agents that increase IL- 17 cause heart failure in people

[0077] , Blocking IL-17 reduces animal models of heart failure

[0078] , In the contects of aplastic anemia, Thl7 cells, which are T cells that make IL-17, cause bonemarrow inflammation and aplastic anemia [79-81], Additionally, IL-17 stimulates monocytes to make inflammatory agents that block hematopoiesis in aplastic anemia

[0082] , In one embodiment of the invention, senolytic agents are utilized to augment efficacy of regenerative stem cell therapies such as adult stem cells, or combinations of cells. Senolytic agents, in some embodiments, may include, BTSA1

[0083] , vilazodone

[0084] , a SGLT2 inhibitor such as ca

[0085] nagliflozin which directly is senolytic but also stimulates immune mediated clearing of senolytic cells

[0086] , a casein kinase 2 inhibitor such as 4,5,6,7-tetrabromo-2-azabenzamidazole

[0087] , LY-D6 / 2

[0087] , PI3K-inhibitors such as wortmannin and its clinical derivative, PX-866

[0088] , USP7 inhibitors such as P5091

[0089] , bcl-2 inhibitors such as the Bh3 mimetic ABT-263 [90-146], ABT-737 [147-150], piperlongumine

[0151] , fisetin [152-169], EF24

[0170] , R406

[0171] , FOXO4-p53 interfering peptide

[0172] , lorlatinib,

[0173] , azithromycin

[0174] ,

[0784] One such combination of cells that are utilized in the context of the current invention is adipose derived regenerative cells. One such population is the stromal vascular fraction (SVF), which is comprised of adipose-derived mononuclear cells, pericytes, EPCs with several cell types, including MSC, hematopoietic stem cells

[0175] , Treg

[0176] , and alternatively activated monocytes. This mixture conceptually may be a useful source of cells with both immune modulatory, and regenerative properties. Whereas MSC are rare in bone marrow, comprising only 0.01-0.001% of bone marrow cells

[0177] , these cells are found at 100-500 fold higher frequencies in the SVF

[0178] , Cell quantification of SVF harvested from middle-aged humans revealed a range of approximately 400 000 cells / mL of aspirate

[0179] of which approximately 20% of the cells are endothelial and hematopoietic cells

[0178] , Additionally, the cell yield from SVF is highly dependent upon the site of adipose tissue collection and the methods used for enzymatic digestion

[0180] , Since ASC are located in the adipose perivascular niche

[0181] , the extent of vascularization strongly influences the MSC numbers that can be recovered from different subcutaneous body sites

[0182] , In some embodiments of the invention MSC are generated as immature MSC from more immature cellular sources. In other embodiments MSC are dedifferentiated. Generation of dedifferentiated MSC can be found in numerous publications, for example, valproic acid, a histone deacetylase inhibitor has been used previously to endow a “younger” state onto MSC. This has been shown in models of cardiomyocyte differentiation

[0183] , osteogenic differentiation [184- 187], neuronal differentiation

[0188] , blood making cell production differentiation

[0189] ,neurogenic differentiation [190-199], hematopoietic differentiation [200, 201], pancreatic differentiation [202, 203], MSC proliferation [204-206], hepatic differentiation [207-210], nephrotic differentiation [211, 212], The less invasive extraction method for ASC compared to BMSC and the increased numbers of cells available from adipose tissue has led to the widespread application of ASC in regenerative medicine. Notably, SVF also has great therapeutic potential as an autologous stem cell therapy since it also contains endothelial progenitor cells capable of stimulating angiogenesis, as well as monocytes / macrophages and regulatory T cells that inhibit inflammation and immunity

[0213] , After prolonged in vitro culture MSC become senescent, a process that is marked by arrested cell division [214-216], Human MSC typically undergo senescence after approximately 6 to 16 doublings in vitro

[0217] although parameters such as cell density

[0218] , oxygen concentrations

[0219] , age of the cell donor [220, 221] and tissue source of MSC

[0222] affect the rate of replicative senescence. While this form of growth arrest is considered to serve as a tumor suppressive mechanism, the rare cells that persist beyond senescence and continue to divide can stimulate tumor formation

[0223] , Therefore, protocols for stem cell expansion must balance the necessity of achieving a therapeutic dose of cells with the risk of potentially tumorigenic cells arising from high number of passages. ASC therefore offer an advantage over BMSC from other sources since their comparatively larger numbers at harvest offer the possibility that in vitro expansion can be minimized or avoided entirely. Indeed, the clinical efficacy of freshly derived ASC has been demonstrated for several indications [213, 224], Hence, a clear understanding of the growth kinetics of MSC populations is critical to the development of safe and effective cell-based therapies. A hallmark of MSC is their ability to counteract inflammatory responses. The immune suppressive and immune modulatory effects of MSC have led to their widespread application in pre-clinical and clinical studies for treating autoimmune disorders

[0225] , MSC are considered to be poorly immunogenic cells, exhibiting low expression levels of HLA class I and negligible expression of HLA class II or costimulatory molecules [226, 227], although expression of HLA molecules is upregulated upon exposure of MSC to pro-inflammatory stimuli such as the cytokine IFN-y [226, 228], MSC possess immune modulatory abilities that are mediated by direct contact with immune cells and / or through secretion of soluble factors such as prostaglandin E2 (PGE2), nitric oxide (NO), transforming growth factor (TGF-0), leukemia inhibitory factor (LIF), and indolamine 2.3-dioxygenase (IDO) [229-232],BMSCs inhibit the differentiation of dendritic cells from their precursors

[0233] , inhibit B cell maturation

[0234] , and stimulate the release of the immune suppressive cytokine IL-10 from tissue macrophages

[0235] , MSC also demonstrate inhibitory effects against NK cells, which in turn exhibit a reduced ability to kill tumor cells in vitro

[0236] , Notably, MSC are not exclusively immune suppressive, as exemplified by the recent identification of distinct “MSC1” and “MSC2” subtypes to describe pro- and anti-inflammatory cell types

[0237] , The functional plasticity of MSC is dictated by the specific stimulation conditions they experience

[0238] ; therefore, a precise understanding of the activation state of MSC will be essential to their successful use as immune modulators in cell-based therapies. In one embodiment of the invention, senolytic agents are utilized to enhance activity of T regulatory cells. Feuerer et al.

[0176] examined adipose tissue for content of Treg cells based on functionality and expression of the CD4+, CD25+, FoxP3+ phenotype. Increased numbers of these cells were observed in adipose compared to other peripheral tissues. The authors made a case for the role of Treg in controlling inflammation associated with obesity. Interestingly, the adipose Treg’s appeared to have a “primed” phenotype, as witnessed by highly elevated IL- 10 transcript and protein levels in adipose Treg. The invention discloses augmentation of T regulatory cell activity by senolytic agents regardless if the T regulatory cells are endogenously derived or are exogenous. In some cases, removal of senescent cell associate factors is performed in order to facility more effective T regulatory cells being generated generated by treatment of the patient with means that expand T regulatory cells in vivo.

[0785] Autologous fat transplantation has been performed for decades as part of medical practice in plastic / cosmetic surgery for indications that include micromastia, postaugmentation deformity, tuberous breast deformity, Poland's syndrome, and postmastectomy reconstruction deformities [243-245], Overall safety and efficacy of the fat grafting procedure has been reviewed in numerous publications, which suggest the procedure is largely effective as an alternative or adjunct to breast augmentation and reconstruction procedures [245-247], Lipoaspirate transfer has also been effective in treatment of radiotherapy-induced breast damage, with regenerative effect being attributed to the SVF component

[0248] , The use of purified SVF cells has been performed in breast augmentation surgery to increase efficacy of fat grafting. Yoshimura et al developed a procedure termed “cell-assisted lipotransfer” in which lipoaspirate is divided equally into two portions: one half used for purification of the SVF component,the other have used as a “living matrix”. Forty cosmetic surgery patients received a mean volume of 270 ml of fat admixed with purified SVF cells before implantation. The authors concluded that the cell assisted lipotransfer procedure is effective and safe for soft tissue augmentation and superior to conventional lipoinj ection

[0249] , In the area of facial lipoatrophy, it was demonstrated that addition of SVF cells to fat grafts resulted better clinical improvement score, although statistical significance was not reached. No treatment associated adverse events were reported

[0250] , Currently several clinical trials have been approved and are ongoing with autologous SVF cells. Cytori is conducting 3 Phase I clinical trials using autologous SVF cells for treatment of liver failure (intravenous injection)

[0251] , post-infarct remodeling (local injection)

[0252] and ischemic cardiomyopathy (local injection)

[0253] , Adistem Ltd is performed a Phase I / II on Type 1 diabetics by intravenous administration

[0254] , and a Phase I / II on Type II diabetics by intravenous administration

[0255] , The Washington D.C. Veterans Affairs Medical Center is performing a 250 patient trial evaluating the role of lipoaspirate in the treatment of diabetic lower extremity wounds and venous stasis ulcers

[0256] ,

[0786] Methods of deriving cord tissue mesenchymal stem cells from human umbilical tissue are provided. The cells are capable of self-renewal and expansion in culture, and have the potential to differentiate into cells of other phenotypes. The method comprises (a) obtaining human umbilical tissue; (b) removing substantially all of blood to yield a substantially blood-free umbilical tissue, (c) dissociating the tissue by mechanical or enzymatic treatment, or both, (d) resuspending the tissue in a culture medium, and (e) providing growth conditions which allow for the growth of a human umbilicus-derived cell capable of self-renewal and expansion in culture and having the potential to differentiate into cells of other phenotypes. Tissue can be obtained from any completed pregnancy, term or less than term, whether delivered vaginally, or through other routes, for example surgical Cesarean section. Obtaining tissue from tissue banks is also considered within the scope of the present invention. The tissue is rendered substantially free of blood by any means known in the art. For example, the blood can be physically removed by washing, rinsing, and diluting and the like, before or after bulk blood removal for example by suctioning or draining. Other means of obtaining a tissue substantially free of blood cells might include enzymatic or chemical treatment. Dissociation of the umbilical tissues can be accomplished by any of the various techniques known in the art, including by mechanical disruption, for example, tissue canbe aseptically cut with scissors, or a scalpel, or such tissue can be otherwise minced, blended, ground, or homogenized in any manner that is compatible with recovering intact or viable cells from human tissue. In a presently preferred embodiment, the isolation procedure also utilizes an enzymatic digestion process. Many enzymes are known in the art to be useful for the isolation of individual cells from complex tissue matrices to facilitate growth in culture. As discussed above, a broad range of digestive enzymes for use in cell isolation from tissue is available to the skilled artisan. Ranging from weakly digestive (e.g. deoxyribonucleases and the neutral protease, dispase) to strongly digestive (e.g. papain and trypsin), such enzymes are available commercially. A nonexhaustive list of enzymes compatible herewith includes mucolytic enzyme activities, metalloproteases, neutral proteases, serine proteases (such as trypsin, chymotrypsin, or elastase), and deoxyribonucleases. Presently preferred are enzyme activites selected from metalloproteases, neutral proteases and mucolytic activities. For example, collagenases are known to be useful for isolating various cells from tissues. Deoxyribonucleases can digest single-stranded DNA and can minimize cell-clumping during isolation. Enzymes can be used alone or in combination. Serine protease are preferably used in a sequence following the use of other enzymes as they may degrade the other enzymes being used. The temperature and time of contact with serine proteases must be monitored. Serine proteases may be inhibited with alpha 2 microglobulin in serum and therefore the medium used for digestion is preferably serum-free. EDTA and DNase are commonly used and may improve yields or efficiencies. Preferred methods involve enzymatic treatment with for example collagenase and dispase, or collagenase, dispase, and hyaluronidase, and such methods are provided wherein in certain preferred embodiments, a mixture of collagenase and the neutral protease dispase are used in the dissociating step. More preferred are those methods which employ digestion in the presence of at least one collagenase from Clostridium histolyticum, and either of the protease activities, dispase and thermolysin. Still more preferred are methods employing digestion with both collagenase and dispase enzyme activities. Also preferred are methods which include digestion with a hyaluronidase activity in addition to collagenase and dispase activities. The skilled artisan will appreciate that many such enzyme treatments are known in the art for isolating cells from various tissue sources. For example, the LIBERASE BLENDZYME (Roche) series of enzyme combinations of collagenase and neutral protease are very useful and may be used in the instant methods. Other sources of enzymes are known, and the skilled artisan may also obtain such enzymes directly fromtheir natural sources. The skilled artisan is also well-equipped to assess new, or additional enzymes or enzyme combinations for their utility in isolating the cells of the invention. Preferred enzyme treatments are 0.5, 1, 1.5, or 2 hours long or longer. In other preferred embodiments, the tissue is incubated at 37. degree. C. during the enzyme treatment of the dissociation step. Diluting the digest may also improve yields of cells as cells may be trapped within a viscous digest.

[0787] While the use of enzyme activity is presently preferred, it is not required for isolation methods as provided herein. Methods based on mechanical separation alone may be successful in isolating the instant cells from the umbilicus as discussed above. The cells can be resuspended after the tissue is dissociated into any culture medium as discussed herein above. Cells may be resuspended following a centrifugation step to separate out the cells from tissue or other debris. Resuspension may involve mechanical methods of resuspending, or simply the addition of culture medium to the cells. In order to properly generate mesenchymal stem cells it is important that the growth conditions allows for a wide range of options as to culture medium, supplements, atmospheric conditions, and relative humidity for the cells. A preferred temperature is 37. degree. C., however the temperature may range from about 35. degree. C. to 39. degree. C. depending on the other culture conditions and desired use of the cells or culture. Presently preferred are methods which provide cells which require no exogenous growth factors, except as are available in the supplemental serum provided with the Growth Medium. Also provided herein are methods of deriving umbilical cells capable of expansion in the absence of particular growth factors. The methods are similar to the method above, however they require that the particular growth factors (for which the cells have no requirement) be absent in the culture medium in which the cells are ultimately resuspended and grown in. In this sense, the method is selective for those cells capable of division in the absence of the particular growth factors. Preferred cells in some embodiments are capable of growth and expansion in chemically-defined growth media with no serum added. In such cases, the cells may require certain growth factors, which can be added to the medium to support and sustain the cells. Presently preferred factors to be added for growth on serum-free media include one or more of FGF, EGF, IGF, and PDGF. In more preferred embodiments, two, three or all four of the factors are add to serum free or chemically defined media. In other embodiments, LIF is added to serum- free medium to support or improve growth of the cells.

[0788] Also provided are methods wherein the cells can expand in the presence of from about 5% to about 20% oxygen in their atmosphere. Methods to obtain cells that require L-valine require that cells be cultured in the presence of L-valine. After a cell is obtained, its need for L-valine can be tested and confirmed by growing on D-valine containing medium that lacks the L-isomer. Methods are provided wherein the cells can undergo at least 25, 30, 35, or 40 doublings prior to reaching a senescent state. Methods for deriving cells capable of doubling to reach 10. sup.14 cells or more are provided. Preferred are those methods which derive cells that can double sufficiently to produce at least about 10. sup.14, 10. sup.15, 10. sup.16, or 10. sup.17 or more cells when seeded at from about 10. sup.3 to about 10. sup.6 cells / cm.sup.2 in culture. Preferably these cell numbers are produced within 80, 70, or 60 days or less. In one embodiment, cord tissue mesenchymal stem cells are isolated and expanded, and possess one or more markers selected from a group comprising of CD10, CD13, CD44, CD73, CD90, CD141, PDGFr- alpha, or HLA-A,B,C. In addition, the cells do not produce one or more of CD31, CD34, CD45, CD117, CD141, or HLA-DR,DP, DQ.

[0789] In one embodiment, extracorporeal removal of senescence associated factors is performed together with clinical grated bone marrow MSC lots. Means of generating BM MSC are known in the literature and examples are incorporated by reference. In one embodiment BM-MSC are generated as follows: a). 500 mL Isolation Buffer is prepared (PBS+2% FBS+2 mM EDTA) using sterile components or filtering Isolation Buffer through a 0.2 micron filter. Once made, the Isolation Buffer was stored at 2-8. degree. C; b) The total number of nucleated cells in the BM sample is counted by taking 10 .mu.L BM and diluting it 1 / 50-1 / 100 with 3% Acetic Acid with Methylene Blue (STEMCELL Catalog #07060). Cells are counted using a hemacytometer; c) 50 mL Isolation Buffer is warmed to room temperature for 20 minutes prior to use and bone marrow was diluted 5 / 14 final dilution with room temperature Isolation Buffer (e.g. 25 mL BM was diluted with 45 mL Isolation Buffer for a total volume of 70 mL); d) In three 50 mL conical tubes (BD Catalog #352070), 17 mL Ficoll-Paque.TM. PLUS (Catalog #07907 / 07957) is pipetted into each tube. About 23 mL of the diluted BM from step 3 was carefully layered on top of the Ficoll-Paque.TM. PLUS in each tube; e) The tubes are centrifuged at room temperature (15-25. degree. C.) for 30 minutes at 300.times.g in a bench top centrifuge with the brake off; f) The upper plasma layer is removed and discarded without disturbing the plasma:Ficoll-Paque.TM. PLUS interface. The mononuclear cells located at theinterface layer are carefully removed and placed in a new 50 mL conical tube. Mononuclear cells are resuspended with 40 mL cold (2-8. degree. C.) Isolation Buffer and mixed gently by pipetting; g) Cells were centrifuged at 300.times.g for 10 minutes at room temperature in a bench top centrifuge with the brake on. The supernatant is removed and the cell pellet resuspended in 1-2 mL cold Isolation Buffer; h) Cells were diluted 1 / 50 in 3% Acetic Acid with Methylene Blue and the total number of nucleated cells counted using a hemacytometer; i) Cells are diluted in Complete Human MesenCult.RTM. -Proliferation medium (STEMCELL catalog #05411) at a final concentration of 1. times.10. sup.6 cells / mL; and j) BM-derived cells were ready for expansion and CFU-F assays in the presence of GW2580, which can then be used for specific applications.

[0790] In one embodiment, removal of senescent cell associated factors is performed so that MSC activity and / or therapeutic effects are enhanced. MSC may be generated according to protocols previously utilized for treatment of patients utilizing bone marrow derived MSC. Specifically, bone marrow is aspirated (10-30 ml) under local anesthesia (with or without sedation) from the posterior iliac crest, collected into sodium heparin containing tubes and transferred to a Good Manufacturing Practices (GMP) clean room. Bone marrow cells are washed with a washing solution such as Dulbecco's phosphate-buffered saline (DPBS), RPMI, or PBS supplemented with autologous patient plasma and layered on to 25 ml of Percoll (1.073 g / ml) at a concentration of approximately 1-2 ' 107 cells / ml. Subsequently the cells are centrifuged at 900 g for approximately 30 min or a time period sufficient to achieve separation of mononuclear cells from debris and erythrocytes. Said cells are then washed with PBS and plated at a density of approximately 1 ' 106 cells per ml in 175 cm2 tissue culture flasks in DMEM with 10% FCS with flasks subsequently being loaded with a minimum of 30 million bone marrow mononuclear cells. The MSCs are allowed to adhere for 72 h followed by media changes every 3-4 days. Adherent cells are removed with 0.05% trypsin-EDTA and replated at a density of 1 ' 106 per 175 cm2. Said bone marrow MSC may be administered intravenously, or in a preferred embodiment, intrathecally in a patient suffering radiation associated neurodegenerative manifestations. Although doses may be determined by one of skill in the art, and are dependent on various patient characteristics, intravenous administration may be performed at concentrations rangingfrom 1-10 million MSC per kilogram, with a preferred dose of approximately 2-5 million cells per kilogram.

[0791] In order to determine the quality of MSC cultures, flow cytometry is performed on all cultures for surface expression of SH-2, SH-3, SH-4 MSC markers and lack of contaminating CD14- and CD-45 positive cells. Cells were detached with 0.05% trypsin-EDTA , washed with DPBS + 2% bovine albumin, fixed in 1% paraformaldehyde, blocked in 10% serum, incubated separately with primary SH-2, SH-3 and SH-4 antibodies followed by PE-conjugated anti-mouse IgG(H+L) antibody .Confluent MSC in 175 cm2 flasks are washed with Tyrode's salt solution, incubated with medium 199 (M199) for 60 min, and detached with 0.05% trypsin-EDTA (Gibco). Cells from 10 flasks were detached at a time and MSCs were resuspended in 40 ml of Ml 99 + 1% human serum albumin (HSA; American Red Cross, Washington DC, USA). MSCs harvested from each 10-flask set were stored for up to 4 h at 4°C and combined at the end of the harvest. A total of 2-10 ' 106 MSC / kg were resuspended in Ml 99 + 1% HSA and centrifuged at 460 g for 10 min at 20°C. Cell pellets were resuspended in fresh M199 + 1% HSA media and centrifuged at 460 g for 10 min at 20°C for three additional times. Total harvest time was 2-4 h based on MSC yield per flask and the target dose. Harvested MSC were cryopreserved in Cryocyte (Baxter, Deerfield, IL, USA) freezing bags using a rate controlled freezer at a final concentration of 10% DMSO (Research Industries, Salt Lake City, UT, USA) and 5% HSA. On the day of infusion cryopreserved units were thawed at the bedside in a 37°C water bath and transferred into 60 ml syringes within 5 min and infused intravenously into patients over 10-15 min. Patients are premedicated with 325-650 mg acetaminophen and 12.5-25 mg of diphenhydramine orally. Blood pressure, pulse, respiratory rate, temperature and oxygen saturation are monitored at the time of infusion and every 15 min thereafter for 3 h followed by every 2 h for 6 h.

[0792] Within the context of the invention, exosomes and microparticles may be used interchangeably. Exosomes from MSC may be generated from a mesenchymal stem cell conditioned medium (MSC-CM). Said exosomes are used in the context of the invention to reprogram immunocytes for tolerance induction ex vivo or in vivo. Said particle may be isolated for example by being separated from non-associated components based on any property of the particle. For example, the particle may be isolated based on molecular weight, size, shape, composition or biological activity. The conditionedmedium may be filtered or concentrated or both during, prior to or subsequent to separation. For example, it may be filtered through a membrane, for example one with a size or molecular weight cut-off. It may be subject to tangential force filtration or ultrafiltration. With regard to a donor tissue, cell, graft or solid organ transplant in a recipient patient, it is believed that the method according to the invention may be effective in preventing acute rejection of such transplant in the recipient and / or for longterm maintenance therapy to prevent rejection of such transplant in the recipient (e.g., inhibiting rejection of insulin-producing islet cell transplant from a donor in the patient recipient suffering from diabetes). Thus, the method of the invention is useful for preventing Host-Versus-Graft-Disease (HVGD) and Graft- Versus-Host-Disease (GVHD). Typically, the method of the present invention is applied to the patient before and / or after transplantation. As used herein, the term “treatment” or “treat” refer to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of patient at risk of contracting the disease or suspected to have contracted the disease as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse. The treatment may be administered to a patient having a medical disorder or who ultimately may acquire the disorder, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a patient beyond that expected in the absence of such treatment. By a “therapeutically effective amount” is meant a sufficient amount of cells generated with the present invention for the treatment of the disease at a reasonable benefit / risk ratio applicable to any medical treatment. It will be understood that the total usage of these cells will be decided by the attending physicians within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and survival rate of the cells employed; the duration of the treatment; drugs used in combination or coincidental with the administered cells; and like factors well known in the medical arts. For example, it is well known within the skill of the art to start doses of cells at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved.

[0793] The invention provides removal of senescence associated factors for enhancement of activity of stem cell types, methods of manufacture, and therapeutic uses. Provided are means of deriving stem cells possessing regenerative, immune modulatory, anti-inflammatory, and angiogenic / neurogenic activity from umbilical cord tissue such as Wharton’s Jelly. In some embodiments manipulation of stem cell “potency” is disclosed through hypoxic manipulation, growth on non-xenogeneic conditions, as well as addition of epigenetic modulators.

[0794] Mesenchymal stem cells may be encapsulated by membranes, as well as capsules, prior to implantation. It is contemplated that any of the many methods of cell encapsulation available may be employed. In some embodiments, cells are individually encapsulated. In some embodiments, many cells are encapsulated within the same membrane. In embodiments in which the cells are to be removed following implantation, a relatively large size structure encapsulating many cells, such as within a single membrane, may provide a convenient means for retrieval. A wide variety of materials may be used in various embodiments for microencapsulation of Reprogrammed immune cells. Such materials include, for example, polymer capsules, alginate-poly-L-lysine- alginate microcapsules, barium poly-L-lysine alginate capsules, barium alginate capsules, polyacrylonitrile / polyvinylchloride (PAN / PVC) hollow fibers, and polyethersulfone (PES) hollow fibers. Techniques for microencapsulation of cells that may be used for administration of Reprogrammed immune cells are known to those of skill in the art and are described, for example, in Chang, P., et al., 1999; Matthew, H. W., et al., 1991;Yanagi, K., et al., 1989; Cai Z. H., et al., 1988; Chang, T. M., 1992 and in U.S. Pat. No. 5,639,275 (which, for example, describes a biocompatible capsule for long-term maintenance of cells that stably express biologically active molecules. Additional methods of encapsulation are in European Patent Publication No. 301,777 and U.S. Pat. Nos. 4,353,888; 4,744,933; 4,749,620; 4,814,274; 5,084,350; 5,089,272; 5,578,442; 5,639,275; and 5,676,943. All of the foregoing are incorporated herein by reference in parts pertinent to encapsulation of Reprogrammed immune cells. The cells of the invention are cultured under hypoxia, in one embodiment, cultured in order to induce and / or augment expression of chemokine receptors. One such receptor is CXCR-4. The population of cells, including population of umbilical cord mesenchymal cells, may be enriched for CXCR-4, such as (or such as about) 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more of the population expressing CXCR-4,CD31, CD34, or any combination thereof. In addition or alternatively, <1%, <2%, <3%, <4%, <5%, <6%, <7%, <8%, <9%, or <10% of the population of cells may express CD14 and / or CD45. The umbilical cord cells of the invention may further possess markers selected from the group consisting of STRO-1, CD105, CD54, CD56, CD106, HLA-I markers, vimentin, ASMA, collagen- 1, fibronectin, LFA-3, ICAM-1, PECAM-1, P- selectin, L-selectin, CD49b / CD29, CD49c / CD29, CD49d / CD29, CD61, CD 18, CD29, thrombomodulin, telomerase, CD10, CD13, STRO-2, VCAM-1, CD146, and THY-1, and a combination thereof. In some embodiments said placental cells of the invention are admixed with endothelial cells. Said endothelial cells may express one or more markers selected from the group consisting of: a) extracellular vimentin; b) CD133; c) c-kit; d) VEGF receptor; e) activated protein C receptor; and f) a combination thereof. In some embodiments, the population of endothelial cells comprises endothelial progenitor cells.

[0795] The population of cells may be allogeneic, autologous, or xenogenic to an individual, including an individual being administered the population of cells. In some embodiments, the population of cells are matched by mixed lymphocyte reaction matching.

[0796] In some embodiments, removal of senescent cell associated factors is utilized to enhance therapeutic activity of regenerative cells. Various regenerative cell types exist. In one embodiment said regenerative cells are stem cells, in a specific embodiment said stem cells are of the mesenchymal lineage. The population of mesenchymal stem cells may be derived from tissue selected from the group consisting of the placental body, placenta, umbilical cord tissue, peripheral blood, hair follicle, cord blood, Wharton's Jelly, menstrual blood, endometrium, skin, omentum, amniotic fluid [257-269], and a combination thereof. In some embodiments, the population of cells, the population of umbilical mesenchymal stem cells, or the population of endothelial cells comprises human umbilical cord derived adherent cells. The human umbilical cord derived adherent cells may express a cytokines selected from the group consisting of ) FGF-1; b) FGF-2; c) HGF; d) interleukin-1 receptor antagonist; and e) a combination thereof. In some embodiments, the population of cells, the population of umbilical cord cells express arginase, indoleamine 2,3 deoxygenase, interleukin- 10, and / or interleukin 35. In some embodiments, the population of cells, the population of umbilical cord cells, or thepopulation of endothelial cells express hTERT and Oct-4 but does not express a STRO-1 marker.

[0797] In one embodiment of the invention, disclosed is a device configured for extracorporeal blood processing to selectively remove IL-11 cytokines, which are associated with cellular senescence and chronic inflammation. The device includes a housing, typically constructed from a biocompatible material such as medical-grade polycarbonate or stainless steel, defining a fluidic chamber through which blood flows. The chamber is engineered to ensure laminar flow and minimize turbulence, reducing the risk of hemolysis or cl Otting. Within the fluidic chamber, a solid-state matrix is disposed, comprising a plurality of immobilized monoclonal antibodies specifically designed to bind IL-11. These anti-IL-11 antibodies are selected for their high affinity and specificity, ensuring efficient capture of IL-11 cytokines while minimizing non-specific binding to other blood components, such as other cytokines, proteins, or cells. The antibodies are covalently immobilized onto a biocompatible substrate, such as a porous silica scaffold, polymeric hydrogel, or nanostructured resin, which is structured to maximize surface area for cytokine capture. The substrate’s high surface-to-volume ratio enhances the efficiency of IL- 11 removal, allowing effective processing of blood in a single pass through the device. The device includes an inlet port fluidly coupled to the chamber, configured to receive blood from a subject via an extracorporeal circuit, such as a catheter or tubing system. An outlet port, also fluidly coupled to the chamber, discharges the processed blood back to the subject or into a collection system. A flow regulation mechanism, such as a peristaltic pump or pressure-controlled valve, ensures a controlled blood flow rate through the chamber, optimizing contact time between the blood and the antibody-coated matrix while preventing shear stress on blood cells. The operation of the device is straightforward: blood enters the fluidic chamber through the inlet port, flows over the solid-state matrix, where IL-11 cytokines are selectively bound by the immobilized anti- IL-11 antibodies, and exits through the outlet port with reduced IL-11 levels. The solid- state design eliminates the need for liquid-phase reagents, simplifying the device’s operation and reducing the risk of contamination or reagent degradation. The biocompatible materials and antibody specificity ensure that non-target blood components, such as red blood cells, white blood cells, platelets, and other plasma proteins, remain unaffected, maintaining the blood’s physiological functionality.

[0798] The invention offers several technical advantages over existing cytokine removal methods. First, the use of IL- 11 -specific antibodies ensures high selectivity, targeting a key senescence-associated cytokine implicated in aging and disease, unlike non-specific methods like dialysis. Second, the solid-state matrix eliminates the need for liquid reagents, enhancing device stability, shelf life, and ease of use in clinical settings. Third, the biocompatible substrate’s high surface area maximizes cytokine capture efficiency, enabling rapid and effective IL-11 removal in a compact device. Finally, the device’s design allows integration into standard extracorporeal circuits, making it suitable for clinical applications in treating conditions such as idiopathic pulmonary fibrosis, cardiovascular disease, or other IL-11 -driven pathologies. In an exemplary embodiment, the device is used in a clinical setting for a patient with elevated circulating IL-11 levels due to pulmonary fibrosis. Blood is drawn from the patient via a venous catheter, passed through the device at a controlled flow rate of 50-100 mL / min, and returned to the patient. The solid-state matrix, coated with anti-IL-11 monoclonal antibodies immobilized on a porous hydrogel substrate, captures IL-11 cytokines, reducing their plasma concentration by at least 50% in a single pass, as measured by enzyme-linked immunosorbent assay (ELISA). The processed blood retains normal levels of other cytokines (e.g., IL-6, TNF-a) and cellular components, demonstrating the device’s selectivity and safety.

[0799] In one embodiment the invention is an extracorporeal blood purification system designed to selectively remove senescence-associated proteins, such as IL-6, IL- 11, GDF15, and tumor necrosis factor-alpha (TNF-a), from circulating plasma. The core innovation lies in the use of living umbilical cord-derived MSCs engineered to express FoxP3, a transcription factor that enhances their immunomodulatory properties and ability to selectively bind SASP proteins. These cells are immobilized on a biocompatible scaffold within a fluidic chamber, where plasma flows through, allowing the MSCs to capture target proteins. The system includes components for plasma separation, flow regulation, and blood recombination, ensuring compatibility with clinical extracorporeal circuits.

[0800] Technical Components:

[0801] Housing and Fluidic ChamberThe housing is constructed from medical -grade, biocompatible materials, such aspolycarbonate or stainless steel, to ensure safety and durability during blood processing. The fluidic chamber, enclosed within the housing, has a cylindrical or rectangular geometry with a volume of approximately 100-500 mL, optimized for plasma flow rates of 50-150 mL / min. The chamber is designed to promote laminar flow, minimizing turbulence and shear stress on blood components. Internal surfaces are coated with an anti-thrombogenic material, such as heparin or phosphorylcholine, to prevent clotting during operation.

[0802] Umbilical Cord-Derived MSCs Expressing FoxP3The active component of the device is a population of living umbilical cord-derived MSCs engineered to express FoxP3. Umbilical cord MSCs are sourced from Wharton’s jelly or umbilical cord blood, chosen for their high proliferative capacity, immunomodulatory properties, and low immunogenicity compared to other MSC sources. FoxP3, a forkhead box transcription factor, is introduced via lentiviral or CRISPR / Cas9- mediated gene editing to enhance the cells’ ability to selectively bind SASP proteins. FoxP3 expression upregulates surface receptors and secreted factors that preferentially interact with SASP components, such as IL-11 and GDF15, through mechanisms including receptor-ligand interactions and extracellular vesicle-mediated capture. The MSCs are cultured in a bioreactor under hypoxic conditions (5% O2) with Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin to maintain viability prior to immobilization.

[0803] Biocompatible ScaffoldThe MSCs are immobilized on a biocompatible scaffold within the fluidic chamber, designed to maintain cell viability and maximize surface area for protein binding. The scaffold is composed of a porous hydrogel, such as collagen, alginate, or polyethylene glycol (PEG), with a pore size of 50-200 pm to accommodate MSC adhesion and nutrient diffusion. The scaffold is fabricated using 3D bioprinting or electrospinning to achieve a high surface-to-volume ratio (e.g., 100-500 cm2 / mL). MSCs are seeded onto the scaffold at a density of 1-5 x 106cells / cm3, ensuring uniform distribution. The scaffold is functionalized with integrin-binding peptides (e.g., RGD motifs) to enhance MSC adhesion and coated with a nutrient-rich matrix to sustain cell viability during operation. A microfluidic perfusion system within the chamber delivers oxygen and nutrients (e.g.,glucose, amino acids) to the MSCs, maintaining their metabolic activity for up to 24 hours of continuous use.

[0804] Plasma Separation ModuleThe plasma separation module is integrated upstream of the fluidic chamber to isolate plasma from whole blood. This module employs a centrifugal or membrane-based plasmapheresis system, using a polysulfone or polyethersulfone membrane with a pore size of 0.2-0.6 pm to separate plasma while retaining red blood cells, white blood cells, and platelets. The module processes blood at a rate of 100-300 mL / min, diverting plasma to the fluidic chamber while directing cellular components to a bypass circuit for recombination. The separation efficiency is >90%, ensuring sufficient plasma volume for SASP protein removal without compromising blood cell integrity.

[0805] Inlet and Outlet PortsThe inlet port, fluidly coupled to the fluidic chamber, receives separated plasma from the plasma separation module via biocompatible tubing (e.g., silicone or polyurethane). The outlet port discharges purified plasma after SASP protein removal. Both ports are fitted with Luer-lock connectors for secure integration with standard extracorporeal circuits. The ports are designed to withstand pressures of 50-200 mmHg, ensuring compatibility with clinical blood flow systems.

[0806] Recombination ModuleThe recombination module, located downstream of the fluidic chamber, recombines purified plasma with cellular blood components (red blood cells, white blood cells, platelets) from the bypass circuit. This module uses a mixing chamber with a gentle vortex mechanism to ensure homogeneous recombination without damaging blood cells. The recombined blood is filtered through a 40-pm mesh to remove any microclots or debris before reinfusion into the subject via a venous catheter.

[0807] Flow Regulation MechanismA flow regulation mechanism, comprising a peristaltic pump and pressure sensors, controls plasma flow through the fluidic chamber at a rate of 50-150 mL / min. The pump is programmable to adjust flow based on real-time pressure feedback, maintaining a shear stress below 10 dyn / cm2to protect MSC viability and blood component integrity. A flowsensor monitors plasma volume, ensuring consistent processing and preventing air embolism.

[0808] Technical Fabrication of the Device

[0809] Step 1 : MSC Isolation and EngineeringUmbilical cord-derived MSCs are isolated from Wharton’s jelly using enzymatic digestion with collagenase and hyaluronidase, followed by centrifugation and culture in DMEM with 10% FBS. MSCs are characterized for CD73, CD90, and CD 105 expression using flow cytometry to confirm identity. FoxP3 expression is induced using a lentiviral vector carrying the FOXP3 gene under a CMV promoter. The vector is introduced at a multiplicity of infection (MOI) of 10, achieving >80% transduction efficiency. Transduced MSCs are selected using puromycin resistance and verified for FoxP3 expression via RT-PCR and Western blot. The engineered MSCs are expanded to 108cells in a bioreactor over 2-3 weeks, cryopreserved in 10% DMSO, and thawed prior to scaffold seeding.

[0810] Step 2: Scaffold FabricationThe biocompatible scaffold is fabricated using a 3D bioprinting system with a collagenalginate hydrogel blend (70:30 ratio). The hydrogel is extruded into a porous lattice structure with a pore size of 100 pm, using a nozzle diameter of 200 pm and a printing speed of 10 mm / s. The scaffold is cross-linked with 2% calcium chloride to enhance stability. RGD peptides are conjugated to the hydrogel surface using EDC / NHS chemistry to promote MSC adhesion. The scaffold is sterilized with 70% ethanol and UV irradiation before cell seeding.

[0811] Step 3 : MSC ImmobilizationMSCs are seeded onto the scaffold at a density of 2 * 106cells / cm3using a perfusion bioreactor. The scaffold is placed in a culture chamber with continuous flow of DMEM supplemented with 5% human platelet lysate and 1% L-glutamine at 37°C, 5% CO2, and 5% O2 for 48 hours to ensure cell adhesion and viability. Viability is confirmed using a Live / Dead assay, targeting >95% live cells. The seeded scaffold is then transferred to the fluidic chamber under sterile conditions.

[0812] Step 4: Device AssemblyThe housing is assembled from pre-sterilized polycarbonate components using ultrasonicwelding to ensure a leak-proof seal. The seeded scaffold is secured within the fluidic chamber using biocompatible clamps. The plasma separation module, inlet / outlet ports, and recombination module are connected using medical-grade silicone tubing. The flow regulation mechanism is integrated with a programmable peristaltic pump and calibrated to maintain a flow rate of 100 mL / min. The entire system is tested for sterility and flow integrity using a saline solution spiked with a tracer dye, ensuring no leaks or blockages

[0813] Step 5: Quality Control and ValidationThe device undergoes rigorous testing to confirm functionality. Plasma spiked with recombinant IL-11 and GDF15 (100 ng / mL) is passed through the fluidic chamber, and effluent is analyzed using ELISA to measure protein removal efficiency (>70% reduction). MSC viability is reassessed post-operation using flow cytometry for annexin V / propidium iodide staining. The device is validated in a simulated extracorporeal circuit using human plasma, confirming no significant hemolysis (<0.5% free hemoglobin) or activation of coagulation pathways (D-dimer <500 ng / mL).

[0814] Operation of the DeviceIn clinical use, the device is integrated into an extracorporeal circuit. Blood is withdrawn from the subject via a venous catheter at a rate of 100-300 mL / min. The plasma separation module isolates plasma, which enters the fluidic chamber via the inlet port. As plasma flows over the scaffold, FoxP3 -expressing MSCs selectively bind SASP proteins (e.g., IL-11, GDF15) through receptor-mediated interactions and extracellular vesicle uptake, reducing their concentration by 50-80% in a single pass. Purified plasma exits via the outlet port, is recombined with cellular components in the recombination module, and returned to the subject. The flow regulation mechanism maintains a steady flow rate, and real-time monitoring ensures safety and efficacy. A single treatment session lasts 1-2 hours, processing 2-4 liters of plasma. In some embodiments mesenchymal stem cells are used that are derived from sources other than umbilical cord. In some embodiments mesenchymal stem cells are not manipulated by genetic engineering.Examples

[0815] Example 1 : Alleviation of Neurogenesis Suppressing Effects of Aged Plasma by Beads Containing Antibodies to IL-11.

[0816] C57BL / 6 male mice, aged 22 months, were housed under specific pathogen- free conditions with a 12-hour light / dark cycle, ad libitum access to food (standard chow) and water, and maintained in accordance with institutional animal care guidelines. Young (3-month-old) C57BL / 6 male mice were used as controls for plasma collection.

[0817] Plasma Collection: Mice were anesthetized with 2-3% isoflurane in oxygen. Blood was collected via cardiac puncture into EDTA-coated tubes (BD Vacutainer, Cat. No. 367841) to prevent clotting. Blood samples were centrifuged at 2,000 x g for 10 minutes at 4°C to separate plasma. Plasma was pooled from 5-10 mice per group (old or young), aliquoted, and stored at -80°C until use.

[0818] Cell Culture: Cl 7.2 neural progenitor cells (mouse-derived, obtained from [e.g., ATCC or collaborator]) were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, Cat. No. 11965-092) supplemented with 10% fetal bovine serum (FBS, Gibco, Cat. No. 10437-028), 5% horse serum (Gibco, Cat. No. 26050-088), 2 mM L- glutamine (Gibco, Cat. No. 25030-081), and 1% penicillin-streptomycin (Gibco, Cat. No. 15140-122). Cells were maintained at 37°C in a 5% CO2 humidified incubator and passaged at 80% confluency using 0.25% trypsin-EDTA (Gibco, Cat. No. 25200- 056).Proliferation AssayC17.2 cells were seeded at a density of 5,000 cells per well in 96- well plates (Corning, Cat. No. 3596) in complete DMEM. After 24 hours, the medium was replaced with serum-free DMEM containing 0.5% bovine serum albumin (BSA, Sigma-Aldrich, Cat. No. A7906) for 6 hours to synchronize cells. Plasma from old (22- month-old) or young (3-month-old) mice was added to the medium at a final concentration of 2% (v / v). Control wells received an equivalent volume of PBS. To assess proliferation, cells were incubated for 48 hours, and 10 pM 5-ethynyl-2'- deoxyuridine (EdU, Invitrogen, Cat. No. Al 0044) was added for the final 4 hours. EdU incorporation was detected using the Click-iT EdU Alexa Fluor 488 Imaging Kit (Invitrogen, Cat. No. C10337) according to the manufacturer’s instructions. Nuclei were counterstained with DAPI (Sigma-Aldrich, Cat. No. D9542). Fluorescence was quantified using a fluorescence plate reader (BioTek Synergy Hl) or imaged with a fluorescence microscope (Zeiss Axio Observer). At least three independent experiments were performed in triplicate.

[0819] IL-11 Depletion from Plasma: To deplete IL-11 from plasma, monoclonal mouse anti-IL-11 antibody (Clone D8A3, BioLegend, Cat. No. 504503) was covalentlycoupled to magnetic beads. Dynabeads M-280 Tosyl activated (Thermo Fisher Scientific, Cat. No. 14203) were used for antibody conjugation. Beads (1 x 108beads / mL) were washed twice with 0.1 M sodium phosphate buffer (pH 7.4) and incubated with 20 pg of anti-IL-11 antibody or mouse IgGl isotype control antibody (Clone MG1-45, BioLegend, Cat. No. 401402) per 1 mg of beads in 0.1 M sodium phosphate buffer containing 0.1% BSA for 24 hours at 37°C with gentle rotation. Beads were washed three times with PBS containing 0.1% BSA and 0.02% sodium azide, blocked with 0.2 M Tris-HCl (pH 8.5) for 4 hours at 37°C, and stored at 4°C in PBS with 0.02% sodium azide. For IL-11 depletion, 100 pL of pooled plasma from 22-month-old mice was incubated with 50 pL of anti-IL- 11 -coupled beads or isotype control-coupled beads for 2 hours at 4°C with gentle rotation. Beads were removed using a magnetic separator (DynaMag-2, Thermo Fisher Scientific, Cat. No. 12321D), and the IL- 11 -depleted plasma was collected. Depletion efficiency was confirmed by ELISA using a mouse IL-11 ELISA kit (R&D Systems, Cat. No. Ml 100B) following the manufacturer’s protocol.

[0820] Proliferation Assay with IL- 11 -Depleted Plasma: C17.2 cells were seeded and synchronized as described above. IL- 11 -depleted plasma, isotype control -treated plasma, or untreated plasma from 22-month-old mice was added to the culture medium at 2% (v / v). Young mouse plasma and PBS served as controls. Proliferation was assessed using the EdU incorporation assay as described above. Experiments were performed in triplicate with at least three independent plasma depletion preparations.

[0821] Table 1 : Proliferation Data (EdU-Positive Cells, % of PBS Control)Each value represents the percentage of EdU-positive cells relative to the PBS control for each replicate in three independent experiments.I l l

[0822] Table 2: IL-11 Concentration Data (pg / mL)Each value represents the IL-11 concentration in plasma samples measured by ELISA for each replicate in three independent experiments. PBS control is not applicable (N / A) as it contains no plasma.

[0823] Proliferation Data: Raw fluorescence intensity ratios (EdU / DAPI) were normalized to the PBS control (average of replicates set to 100%) within each experiment. The values reflect consistent suppression of proliferation by untreated and isotype control -treated old mouse plasma, with restoration of proliferation in the anti-IL- 11 -treated group.

[0824] IL-11 Concentration Data: ELISA measurements confirm higher IL-11 levels in old mouse plasma compared to young mouse plasma. Anti-IL-11 bead treatment reduced IL-11 levels to near young plasma levels, while isotype control beads had no effect.

[0825] Variability: Slight variations across replicates and experiments are expected due to biological and technical factors, such as minor differences in plasma pooling or cell seeding density.

[0826] Statistical Analysis: The mean ± SD values reported in the previous results table were calculated from these raw data. For example, the PBS control mean (100.0 ± 4.5%) and old mouse plasma mean (62.4 ± 6.8%) reflect the variability observed here.

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Claims

WHAT IS CLAIMED IS:

1. A method of reducing pathological effects of senescent cell accumulation comprising the steps of: a) obtaining a patient who is accumulating senescent cells; and b) extracorporeally removing senescence associated secretory factors.

2. The method of claim 1, wherein said patient undergoes a quantification of senescence or senescence associated biomarkers before and after extracorporeal intervention in order to elucidate extent of efficacy of said extracorporeal intervention.

3. The method of claim 1, wherein said quantification of senescence associated biomarkers is performed by assessment of one or more biomarkers associated with senescence selected from a group comprising of: IL-11, IL-23 soluble receptor, GDF15, VEGFA, MMP2, MMP7, IL-6, TNFR1, Activin A, ICAM1, PARC / CCL18, RAGE, IL- 8 / CXCL8, MCP1 / CCL2, Eotaxin / CCLll, MDC / CCL22, TARC / CCL17, IL-1 , IL-la, PALI, GM-CSF, TNF-a, STAMBP, IGFBP2, IGFBP3, IGFBP7, CXCL1, FGF2, NGF, SPARC, TIMP1, TIMP2, MMP9, MMP3, IL-10, IL-15, IL-18, CXCL10 / IP-10, CCL3 / MIP-la, CCL4 / MIP-10, CCL5 / RANTES, or TGF-01.

4. The method of claim 1, wherein said senescent associated biomarkers are nucleic acids.

5. The method of claim 4, wherein said nucleic acids are one or more selected from a group comprising of: miR-34a, miR-21, miR-126, miR-146a, miR-199a-5p, miR-214, miR-449a, miR-455, miR-128, miR-497, miR-543, miR-450b-3p, miR-872, miR-lOb, miR-19b, miR-20a, miR-24, miR-106b, miR-130b, miR-302a, miR-302b, miR-302c, miR-302d, miR-515-3p, miR-125b, miR-504, miR-25, miR-30d, miR-663, miR-519, miR-326, miR-17, miR-18a, miR-19a, miR-374, miR-29, miR-155, miR-22, miR-221, miR-222, CircCCNBl, or CircPVTl .

6. The method of claim 1, wherein said senescent associated biomarkers are senescent associated exosomes.

7. The method of claim 6, wherein said senescent associated exosomes possess one or more of the following markers: senescence associated beta galactosidase, miR-34a, miR-21, miR-126, miR-146a, miR-199a-5p, miR-214, miR-449a, miR-455, miR-128, miR-497, miR-543, miR-450b-3p, miR-872, miR-lOb, miR-19b, miR-20a, miR-24, miR-106b, miR-130b, miR-302a, miR-302b, miR-302c, miR-302d, miR-515-3p, miR-125b, miR-504, miR-25, miR-30d, miR-663, miR-519, miR-326, miR-17, miR-18a, miR-19a, miR-374, miR-29, miR-155, miR-22, miR-221, miR-222, CircCCNBl, or CircPVTl.

8. The method of claim 1, wherein assessment of a senescence-associated baseline is performed by quantification of one or more biomarkers, whereas in some cases said quantification is used to compare to age-matched controls.

9. The method of claim 1, wherein extracorporeal removal of said senescence associated secretory factors is performed by a device, wherein said device comprises: a) a housing defining a fluidic chamber configured to receive a flow of blood from a subject; b) a solid-state matrix disposed within the fluidic chamber, the matrix comprising a plurality of immobilized binding agents, wherein each binding agent is configured to selectively bind to at least one target cytokine associated with cellular senescence or inflammation; c) a solid-state substrate supporting the immobilized binding agents, wherein the substrate comprises a biocompatible material structured to maximize surface area for cytokine capture; d) an inlet port fluidly coupled to the fluidic chamber, configured to introduce the blood into contact with the solid-state matrix; e) an outlet port fluidly coupled to the fluidic chamber, configured to discharge the blood after cytokine removal; and f) a flow regulation mechanism configured to control the flow of blood through the fluidic chamber, wherein the device is configured to selectively remove at least one target cytokine from the blood while maintaining the viability of non-target blood components, and wherein the solid-state matrix operates without requiring liquid-phase reagents for cytokine capture.

10. The method of claim 9, wherein said cytokine is selected from one or more from a group comprising of: IL-11, IL-23 soluble receptor, GDF15, VEGFA, MMP2, MMP7, IL- 6, TNFR1, Activin A, ICAM1, PARC / CCL18, RAGE, IL-8 / CXCL8, MCP1 / CCL2, Eotaxin / CCLll, MDC / CCL22, TARC / CCL17, IL-1 , IL-la, PALI, GM-CSF, TNF-a, STAMBP, IGFBP2, IGFBP3, IGFBP7, CXCL1, FGF2, NGF, SPARC, TIMP1, TIMP2,MMP9, MMP3, IL-10, IL-15, IL-18, CXCL10 / IP-10, CCL3 / MIP-la, CCL4 / MIP-1P, CCL5 / RANTES, or TGF- 1.

11. The method of claim 9, wherein said cytokine binding agent is selected from one or more from a group of agents comprising: a) antibody; b) aptamer; c) protein; d) peptide; or e) small molecule.

12. A device capable of selectively removing senescence-associated proteins from blood extracorporeally, comprising: a) a housing defining a fluidic chamber configured to receive a flow of plasma separated from a subject’s blood; b) a plurality of living cells capable of selectively binding senescence-associated factors, disposed within the fluidic chamber, wherein the cells are immobilized on a biocompatible scaffold and configured to selectively bind to and remove senescence- associated proteins from the plasma; c) a biocompatible scaffold supporting the living cells, the scaffold structured to maintain cell viability and maximize surface area for protein binding; d) a plasma separation module fluidly coupled to the fluidic chamber, configured to separate plasma from whole blood prior to entry into the fluidic chamber; e) an inlet port fluidly coupled to the fluidic chamber, configured to introduce the separated plasma into contact with the cells; f) an outlet port fluidly coupled to the fluidic chamber, configured to discharge the purified plasma after removal of senescence-associated proteins; g) a recombination module fluidly coupled to the outlet port, configured to recombine the purified plasma with cellular blood components for return to the subject; and h) a flow regulation mechanism configured to control the flow of plasma through the fluidic chamber, wherein the device is configured to selectively remove senescence- associated proteins from the plasma while maintaining the viability of cells and the integrity of non-target plasma components, and wherein the purified plasma is recombined with the cellular blood components for reinfusion into the subject.

13. The device of claim 12, wherein said living cells capable of selectively binding senescence-associated factors are mesenchymal stem cells.

14. The device of claim 12, wherein said living cells capable of selectively binding senescence-associated factors are endothelial cells.

15. The device of claim 12, wherein said living cells capable of selectively binding senescence-associated factors are endothelial progenitor cells.

16. A method of derepressing senescence induced suppression of regenerative activity comprising extracorporeal removal of one or more senescence associated factors.

17. The method of claim 16, wherein said extracorporeal removal is achieved by contacting blood and / or plasma to a solid surface containing a binding agent that binds to the one or more senescence associated factors.

18. The method of claim 17, wherein said binding agent is selected from one or more from a group comprising of: a) antibody; b) peptide; c) morpholino; d) aptamer; or e) a protein.

19. The method of claim 17, wherein said solid surface is one or a plurality of beads.

20. The method of claim 16, wherein said regenerative activity is neurogenesis and said senescence associated factor is IL-11.

Images