Treatment of infertility with extracellular vesicle composition

Abstract

Disclosed are methods of treating infertility or increasing fertility of a female subject by administering a therapeutic MSC secretome product made by a method comprising culturing bone marrow-derived MSCs under conditions that include oxygen tension below 5% and a culture media with a pH below 7.

Description

Attorney Docket No.66309-727.601 TREATMENT OF INFERTILITY WITH EXTRACELLULAR VESICLE COMPOSITION CROSS REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 563,042 filedon March 8, 2024, the entirety of which is hereby incorporated by reference herein. BACKGROUND

[0002] Infertility is a significant health concern for many people around the globe, and theglobal disease burden has been rising for decades. Female infertility can result from genetic conditions, gonadotoxic chemotherapeutics, environmental and lifestyle exposures (e.g., smoking, pollutants), natural aging processes and iatrogenic causes. Considering the different causes of female infertility, clinical research into therapeutic candidates that employ novel mechanisms of action is important to the development of more effective solutions. There exists a need for safe and effective treatments for infertility. SUMMARY

[0003] This Summary introduces a selection of concepts that are described further below in theDetailed Description. This Summary is not intended to limit the scope of the claimed subject matter.

[0004] Disclosed herein are methods, systems, and compositions for treating infertility orsymptoms thereof in a subject. In some embodiments, a method disclosed herein comprises administering to the subject a composition comprising one or more extracellular vesicles (EVs). In some embodiments, the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222- 3p, hsa-miR-27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR- 199a-3p, hsa-miR-221-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR- 423-3p, hsa-miR-424-5p, or hsa-miR-940, or a combination of two or more thereof. In some embodiments, the composition comprises TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocyte-macrophage colony-stimulating factor receptor subunit alpha), GPR115Attorney Docket No.66309-727.601 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein- 3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein- glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), Transferrin, or a combination of two or more thereof. In some embodiments, the subject has a history of infertility. In some embodiments, the subject is from 21 to 50 years of age. In some embodiments, the subject is female. In some embodiments, the subject has a history of one or more failed in vitro fertilization (IVF) cycles. In some embodiments, the subject has a lower level of anti-Mullerian hormone as compared to a subject without a history of infertility. In some embodiments, the subject has premature ovarian failure (POF). In some embodiments, the subject has polycystic ovary syndrome (PCOS). In some embodiments, the subject has one or more of the following symptoms selected from the group consisting of: endometriosis, a thyroid disorder, a blocked fallopian tube, a damaged fallopian tube, pelvic inflammatory disease (PID), a high follicle-stimulating hormone (FSH) level, a low estrogen level, a fibroid, a polyp, an autoimmune disorder, a congenital uterine abnormality, a sexually transmitted infection (STI), a hormonal imbalance, a prolactin imbalance, a cortisol imbalance, a gonadotropin-releasing hormone (GnRH) deficiency, inability to conceive, irregular menstrual periods, absent menstrual periods, painful menstrual periods, cramping, heavy or prolonged menstrual bleeding, endometriosis, hormonal fluctuations, mood disturbances, sudden fluctuations in weight, sleep disturbances, and fatigue, or any combination thereof. In some embodiments, the subject is not menopausal or post-menopausal. In some embodiments, the one or more EVs comprise hsa- miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR- 145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR- 320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, or hsa-miR-940, or a combination of two or more thereof. In some embodiments, the administering comprises intravenous administration. In some embodiments, the intravenous administration is carried out over the course of 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments, the composition is prepared by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-Attorney Docket No.66309-727.601 MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, a method disclosed herein comprises preparing the composition prior to the administering, wherein the preparing the composition occurs by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, wherein the culture media is serum-free. In some embodiments, the culture media has a glucose concentration below 4.5 g / L. In some embodiments, formulating the MSC conditioned media comprises exchanging the conditioned media for a pharmaceutically acceptable formulation. In some embodiments, the pharmaceutically acceptable formulation comprises saline. In some embodiments, the composition comprises at least 6x1010to 8x1010extracellular vesicles per mL and is administered at a dose of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml. In some embodiments, the dose is administered in combination with normal saline at a final volume of 100 mL. In some embodiments, the composition comprises saline (0.9% sodium chloride). In some embodiments, the saline is present in the composition at about 80% to about 95% saline. In some embodiments, the composition comprises sodium chloride, sodium lactate, potassium chloride, and calcium chloride. In some embodiments, the molecular weight of any non- excipient component of the composition is greater than about 10 kDa (kilodaltons). In some embodiments, the composition comprises a polysaccharide. In some embodiments, the polysaccharide is present in the composition at about 0.3 M or about 60 mM. In some embodiments, any non-excipient component of the composition has a size of less than about 0.2 microns. In some embodiments, the composition is sterile by USP <71>. In some embodiments, the composition is endotoxin USP <85> free. In some embodiments, the composition is negative for mycoplasma DNA. In some embodiments, the composition is cell-free. In some embodiments, the composition is stored between -80 °C and -60 °C. In some embodiments, the composition is administered within 6 hours of thaw when maintained at ambient temperature. In some embodiments, the composition is present in a glass vial. In some embodiments, the composition is formulated for intravenous administration. In some embodiments, the composition has a pH of about 6 to about 7.5. In some embodiments, the BM-MSCs are negative for CD14, CD31, CD34, and CD45. In some embodiments, the BM-MSCs are positive for CD73, CD105, CD166, and CD90. In some embodiments, the BM-MSCs are capable of undergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyteAttorney Docket No.66309-727.601 phenotypes. In some embodiments, the BM-MSCs are obtained from an iliac crest aspiration of a single donor. In some embodiments, a concentration of the one or more EVs in the composition is measured by nanoparticle tracking analysis (NTA). In some embodiments, the NTA comprises light scatter and fluorescence evaluation. In some embodiments, the concentration of the one or more EVs is about 10 billion to about 250 billion EVs per mL of the composition. In some embodiments, the concentration of the one or more EVs is at about 1 billion to about 40 billion EVs per ml of the composition. In some embodiments, the one or more EVs have an average diameter of about 30 nm to about 170 nm. In some embodiments, a total protein concentration of the composition is about 10 to about 40 µg per ml of the composition. In some embodiments, a total protein concentration of the composition is about 1.5 to about 6 µg per ml of the composition. In some embodiments, the total protein concentration is measured by ELISA. In some embodiments, the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, and Let-7. In some embodiments, the one or more EVs comprise TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), and MIF (Macrophage migration inhibitory factor).

[0005] Disclosed herein are methods and compositions for treating infertility or symptomsthereof in a subject. In some embodiments, a method disclosed herein comprises administering to the subject a composition comprising one or more extracellular vesicles (EVs). In some embodiments, at least 80% of the EVs are CD63+, CD9-, CD81-. In some embodiments, the subject has a history of infertility. In some embodiments, the subject is from 21 to 50 years of age. In some embodiments, the subject is female. In some embodiments, the subject has a history of one or more failed in vitro fertilization (IVF) cycles. In some embodiments, the subject has a lower level of anti-Mullerian hormone as compared to a subject without a history of infertility. In some embodiments, the subject has premature ovarian failure (POF). In some embodiments, the subject has polycystic ovary syndrome (PCOS). In some embodiments, the subject has one or more of the following symptoms selected from the group consisting of: endometriosis, a thyroid disorder, a blocked fallopian tube, a damaged fallopian tube, pelvic inflammatory disease (PID), a high follicle-stimulating hormone (FSH) level, a low estrogen level, a fibroid, a polyp, an autoimmune disorder, a congenital uterine abnormality, a sexually transmitted infection (STI), a hormonal imbalance, a prolactin imbalance, a cortisol imbalance, a gonadotropin-releasing hormone (GnRH) deficiency, inability to conceive, irregular menstrual periods, absent menstrual periods, painful menstrual periods, cramping, heavy or prolonged menstrual bleeding, endometriosis, hormonal fluctuations, mood disturbances, sudden fluctuations in weight, sleep disturbances, and fatigue, or any combination thereof. In someAttorney Docket No.66309-727.601 embodiments, the subject is not menopausal or post-menopausal. In some embodiments, the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa- miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-29a-3p, hsa-miR-29c- 3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, or hsa- miR-940, or a combination of two or more thereof. In some embodiments, the composition comprises TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), bIG-H3 (Transforming growth factor- beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocyte-macrophage colony- stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein-3), IGFBP-6 (Insulin-like growth factor binding protein- 6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein-glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), Transferrin, or a combination of two or more thereof. In some embodiments, the administering comprises intravenous administration. In some embodiments, the intravenous administration is carried out over the course of 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments, the composition is prepared by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, a method disclosed herein comprises preparing the composition prior to the administering, wherein the preparing the composition occurs by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSCAttorney Docket No.66309-727.601 conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, wherein the culture media is serum-free. In some embodiments, the culture media has a glucose concentration below 4.5 g / L. In some embodiments, formulating the MSC conditioned media comprises exchanging the conditioned media for a pharmaceutically acceptable formulation. In some embodiments, the pharmaceutically acceptable formulation comprises saline. In some embodiments, the composition comprises at least 6x1010to 8x1010extracellular vesicles per mL and is administered at a dose of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml. In some embodiments, the dose is administered in combination with normal saline at a final volume of 100 mL. In some embodiments, the composition comprises saline (0.9% sodium chloride). In some embodiments, the saline is present in the composition at about 80% to about 95% saline. In some embodiments, the composition comprises sodium chloride, sodium lactate, potassium chloride, and calcium chloride. In some embodiments, the molecular weight of any non-excipient component of the composition is greater than about 10 kDa (kilodaltons). In some embodiments, the composition comprises a polysaccharide. In some embodiments, the polysaccharide is present in the composition at about 0.3 M or about 60 mM. In some embodiments, any non-excipient component of the composition has a size of less than about 0.2 microns. In some embodiments, the composition is sterile by USP <71>. In some embodiments, the composition is endotoxin USP <85> free. In some embodiments, the composition is negative for mycoplasma DNA. In some embodiments, the composition is cell- free. In some embodiments, the composition is stored between -80 °C and -60 °C. In some embodiments, the composition is administered within 6 hours of thaw when maintained at ambient temperature. In some embodiments, the composition is present in a glass vial. In some embodiments, the composition is formulated for intravenous administration. In some embodiments, the composition has a pH of about 6 to about 7.5. In some embodiments, the BM- MSCs are negative for CD14, CD31, CD34, and CD45. In some embodiments, the BM-MSCs are positive for CD73, CD105, CD166, and CD90. In some embodiments, the BM-MSCs are capable of undergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, the BM-MSCs are obtained from an iliac crest aspiration of a single donor. In some embodiments, a concentration of the one or more EVs in the composition is measured by nanoparticle tracking analysis (NTA). In some embodiments, the NTA comprises light scatter and fluorescence evaluation. In some embodiments, the concentration of the one or more EVs is about 10 billion to about 250 billion EVs per mL of the composition. In some embodiments, the concentration of the one or more EVs is at about 1 billion to about 40 billion EVs per ml of the composition. In some embodiments, the one or more EVs have an average diameter of about 30 nm to about 170 nm. In some embodiments, aAttorney Docket No.66309-727.601 total protein concentration of the composition is about 10 to about 40 µg per ml of the composition. In some embodiments, a total protein concentration of the composition is about 1.5 to about 6 µg per ml of the composition. In some embodiments, the total protein concentration is measured by ELISA. In some embodiments, the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, and Let-7. In some embodiments, the one or more EVs comprise TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), and MIF (Macrophage migration inhibitory factor).

[0006] Disclosed herein are methods and compositions for treating infertility or symptomsthereof in a subject. In some embodiments, a method disclosed herein comprises administering to the subject a composition comprising: TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocyte-macrophage colony-stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein- 3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein- glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), Transferrin, or a combination of two or more thereof. In some embodiments, the subject has a history of infertility. In some embodiments, the subject is from 21 to 50 years of age. In some embodiments, the subject is female. In some embodiments, the subject has a history of one or more failed in vitro fertilization (IVF) cycles. In some embodiments, the subject has a lower level of anti-Mullerian hormone as compared to a subject without a history of infertility. In some embodiments, the subject has premature ovarian failure (POF). In some embodiments, the subject has polycystic ovary syndrome (PCOS). In some embodiments, the subject has one orAttorney Docket No.66309-727.601 more of the following symptoms selected from the group consisting of: endometriosis, a thyroid disorder, a blocked fallopian tube, a damaged fallopian tube, pelvic inflammatory disease (PID), a high follicle-stimulating hormone (FSH) level, a low estrogen level, a fibroid, a polyp, an autoimmune disorder, a congenital uterine abnormality, a sexually transmitted infection (STI), a hormonal imbalance, a prolactin imbalance, a cortisol imbalance, a gonadotropin-releasing hormone (GnRH) deficiency, inability to conceive, irregular menstrual periods, absent menstrual periods, painful menstrual periods, cramping, heavy or prolonged menstrual bleeding, endometriosis, hormonal fluctuations, mood disturbances, sudden fluctuations in weight, sleep disturbances, and fatigue, or any combination thereof. In some embodiments, the subject is not menopausal or post-menopausal. In some embodiments, the one or more EVs comprise hsa- miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR- 145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR- 320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, or hsa-miR-940, or a combination of two or more thereof. In some embodiments, the administering comprises intravenous administration. In some embodiments, the intravenous administration is carried out over the course of 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments, the composition is prepared by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM- MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, a method disclosed herein comprises preparing the composition prior to the administering, wherein the preparing the composition occurs by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, wherein the culture media is serum-free. In some embodiments, the culture media has a glucose concentration below 4.5 g / L. In some embodiments, formulating the MSC conditioned media comprises exchanging the conditioned media for a pharmaceutically acceptable formulation. In some embodiments, the pharmaceutically acceptable formulation comprises saline. In some embodiments, the composition comprises at least 6x1010to 8x1010extracellular vesicles per mL and is administered at a dose of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml. In some embodiments, the dose is administered in combination with normal saline at a final volume of 100 mL. In some embodiments, the composition comprises saline (0.9% sodium chloride). InAttorney Docket No.66309-727.601 some embodiments, the saline is present in the composition at about 80% to about 95% saline. In some embodiments, the composition comprises sodium chloride, sodium lactate, potassium chloride, and calcium chloride. In some embodiments, the molecular weight of any non- excipient component of the composition is greater than about 10 kDa (kilodaltons). In some embodiments, the composition comprises a polysaccharide. In some embodiments, the polysaccharide is present in the composition at about 0.3 M or about 60 mM. In some embodiments, any non-excipient component of the composition has a size of less than about 0.2 microns. In some embodiments, the composition is sterile by USP <71>. In some embodiments, the composition is endotoxin USP <85> free. In some embodiments, the composition is negative for mycoplasma DNA. In some embodiments, the composition is cell-free. In some embodiments, the composition is stored between -80 °C and -60 °C. In some embodiments, the composition is administered within 6 hours of thaw when maintained at ambient temperature. In some embodiments, the composition is present in a glass vial. In some embodiments, the composition is formulated for intravenous administration. In some embodiments, the composition has a pH of about 6 to about 7.5. In some embodiments, the BM-MSCs are negative for CD14, CD31, CD34, and CD45. In some embodiments, the BM-MSCs are positive for CD73, CD105, CD166, and CD90. In some embodiments, the BM-MSCs are capable of undergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, the BM-MSCs are obtained from an iliac crest aspiration of a single donor. In some embodiments, a concentration of the one or more EVs in the composition is measured by nanoparticle tracking analysis (NTA). In some embodiments, the NTA comprises light scatter and fluorescence evaluation. In some embodiments, the concentration of the one or more EVs is about 10 billion to about 250 billion EVs per mL of the composition. In some embodiments, the concentration of the one or more EVs is at about 1 billion to about 40 billion EVs per ml of the composition. In some embodiments, the one or more EVs have an average diameter of about 30 nm to about 170 nm. In some embodiments, a total protein concentration of the composition is about 10 to about 40 µg per ml of the composition. In some embodiments, a total protein concentration of the composition is about 1.5 to about 6 µg per ml of the composition. In some embodiments, the total protein concentration is measured by ELISA. In some embodiments, the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, and Let-7. In some embodiments, the one or more EVs comprise TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), and MIF (Macrophage migration inhibitory factor).Attorney Docket No.66309-727.601

[0007] Disclosed herein are methods and compositions for increasing a level of anti-Mullerianhormone in a subject. In some embodiments, a method disclosed herein comprises administering to the subject a composition comprising one or more extracellular vesicles (EVs). In some embodiments, at least 80% of the EVs are CD63+, CD9-, CD81-. In some embodiments, the subject has a history of infertility. In some embodiments, the subject is from 21 to 50 years of age. In some embodiments, the subject is female. In some embodiments, the subject has a history of one or more failed in vitro fertilization (IVF) cycles. In some embodiments, the subject has a lower level of anti-Mullerian hormone as compared to a subject without a history of infertility. In some embodiments, the subject has premature ovarian failure (POF). In some embodiments, the subject has polycystic ovary syndrome (PCOS). In some embodiments, the subject has one or more of the following symptoms selected from the group consisting of: endometriosis, a thyroid disorder, a blocked fallopian tube, a damaged fallopian tube, pelvic inflammatory disease (PID), a high follicle-stimulating hormone (FSH) level, a low estrogen level, a fibroid, a polyp, an autoimmune disorder, a congenital uterine abnormality, a sexually transmitted infection (STI), a hormonal imbalance, a prolactin imbalance, a cortisol imbalance, a gonadotropin-releasing hormone (GnRH) deficiency, inability to conceive, irregular menstrual periods, absent menstrual periods, painful menstrual periods, cramping, heavy or prolonged menstrual bleeding, endometriosis, hormonal fluctuations, mood disturbances, sudden fluctuations in weight, sleep disturbances, and fatigue, or any combination thereof. In some embodiments, the subject is not menopausal or post-menopausal. In some embodiments, the administering comprises intravenous administration. In some embodiments, the intravenous administration is carried out over the course of 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments, the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa- miR-27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR- 29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa- miR-424-5p, or hsa-miR-940, or a combination of two or more thereof. In some embodiments, the composition comprises TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf- related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocyte- macrophage colony-stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-Attorney Docket No.66309-727.601 coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein-3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein-glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), Transferrin, or a combination of two or more thereof. In some embodiments, the administering comprises intravenous administration. In some embodiments, the intravenous administration is carried out over the course of 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments, the composition is prepared by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, a method disclosed herein comprises preparing the composition prior to the administering, wherein the preparing the composition occurs by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, wherein the culture media is serum-free. In some embodiments, the culture media has a glucose concentration below 4.5 g / L. In some embodiments, formulating the MSC conditioned media comprises exchanging the conditioned media for a pharmaceutically acceptable formulation. In some embodiments, the pharmaceutically acceptable formulation comprises saline. In some embodiments, the composition comprises at least 6x1010to 8x1010extracellular vesicles per mL and is administered at a dose of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml. In some embodiments, the dose is administered in combination with normal saline at a final volume of 100 mL. In some embodiments, the composition comprises saline (0.9% sodium chloride). In some embodiments, the saline is present in the composition at about 80% to about 95% saline. In some embodiments, the composition comprises sodium chloride, sodium lactate, potassium chloride, and calcium chloride. In some embodiments, the molecular weight of any non-excipient component of the composition is greater than about 10 kDa (kilodaltons). In some embodiments, the composition comprises a polysaccharide. In someAttorney Docket No.66309-727.601 embodiments, the polysaccharide is present in the composition at about 0.3 M or about 60 mM. In some embodiments, any non-excipient component of the composition has a size of less than about 0.2 microns. In some embodiments, the composition is sterile by USP <71>. In some embodiments, the composition is endotoxin USP <85> free. In some embodiments, the composition is negative for mycoplasma DNA. In some embodiments, the composition is cell- free. In some embodiments, the composition is stored between -80 °C and -60 °C. In some embodiments, the composition is administered within 6 hours of thaw when maintained at ambient temperature. In some embodiments, the composition is present in a glass vial. In some embodiments, the composition is formulated for intravenous administration. In some embodiments, the composition has a pH of about 6 to about 7.5. In some embodiments, the BM- MSCs are negative for CD14, CD31, CD34, and CD45. In some embodiments, the BM-MSCs are positive for CD73, CD105, CD166, and CD90. In some embodiments, the BM-MSCs are capable of undergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, the BM-MSCs are obtained from an iliac crest aspiration of a single donor. In some embodiments, a concentration of the one or more EVs in the composition is measured by nanoparticle tracking analysis (NTA). In some embodiments, the NTA comprises light scatter and fluorescence evaluation. In some embodiments, the concentration of the one or more EVs is about 10 billion to about 250 billion EVs per mL of the composition. In some embodiments, the concentration of the one or more EVs is at about 1 billion to about 40 billion EVs per ml of the composition. In some embodiments, the one or more EVs have an average diameter of about 30 nm to about 170 nm. In some embodiments, a total protein concentration of the composition is about 10 to about 40 µg per ml of the composition. In some embodiments, a total protein concentration of the composition is about 1.5 to about 6 µg per ml of the composition. In some embodiments, the total protein concentration is measured by ELISA. In some embodiments, the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, and Let-7. In some embodiments, the one or more EVs comprise TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), and MIF (Macrophage migration inhibitory factor).

[0008] Disclosed herein are methods and compositions for increasing a level of anti-Mullerianhormone in a subject. In some embodiments, a method disclosed herein comprises administering to the subject a composition comprising one or more extracellular vesicles (EVs). In some embodiments, the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa- miR-27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-Attorney Docket No.66309-727.601 29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa- miR-424-5p, or hsa-miR-940, or a combination of two or more thereof. In some embodiments, the subject has a history of infertility. In some embodiments, the subject is from 21 to 50 years of age. In some embodiments, the subject is female. In some embodiments, the subject has a history of one or more failed in vitro fertilization (IVF) cycles. In some embodiments, the subject has a lower level of anti-Mullerian hormone as compared to a subject without a history of infertility. In some embodiments, the subject has premature ovarian failure (POF). In some embodiments, the subject has polycystic ovary syndrome (PCOS). In some embodiments, the subject has one or more of the following symptoms selected from the group consisting of: endometriosis, a thyroid disorder, a blocked fallopian tube, a damaged fallopian tube, pelvic inflammatory disease (PID), a high follicle-stimulating hormone (FSH) level, a low estrogen level, a fibroid, a polyp, an autoimmune disorder, a congenital uterine abnormality, a sexually transmitted infection (STI), a hormonal imbalance, a prolactin imbalance, a cortisol imbalance, a gonadotropin-releasing hormone (GnRH) deficiency, inability to conceive, irregular menstrual periods, absent menstrual periods, painful menstrual periods, cramping, heavy or prolonged menstrual bleeding, endometriosis, hormonal fluctuations, mood disturbances, sudden fluctuations in weight, sleep disturbances, and fatigue, or any combination thereof. In some embodiments, the subject is not menopausal or post-menopausal. In some embodiments, the administering comprises intravenous administration. In some embodiments, the intravenous administration is carried out over the course of 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments, the composition comprises TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocyte-macrophage colony-stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein- 3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4Attorney Docket No.66309-727.601 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein- glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), Transferrin, or a combination of two or more thereof. In some embodiments, the administering comprises intravenous administration. In some embodiments, the intravenous administration is carried out over the course of 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments, the composition is prepared by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, a method disclosed herein comprises preparing the composition prior to the administering, wherein the preparing the composition occurs by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, wherein the culture media is serum-free. In some embodiments, the culture media has a glucose concentration below 4.5 g / L. In some embodiments, formulating the MSC conditioned media comprises exchanging the conditioned media for a pharmaceutically acceptable formulation. In some embodiments, the pharmaceutically acceptable formulation comprises saline. In some embodiments, the composition comprises at least 6x1010to 8x1010extracellular vesicles per mL and is administered at a dose of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml. In some embodiments, the dose is administered in combination with normal saline at a final volume of 100 mL. In some embodiments, the composition comprises saline (0.9% sodium chloride). In some embodiments, the saline is present in the composition at about 80% to about 95% saline. In some embodiments, the composition comprises sodium chloride, sodium lactate, potassium chloride, and calcium chloride. In some embodiments, the molecular weight of any non- excipient component of the composition is greater than about 10 kDa (kilodaltons). In some embodiments, the composition comprises a polysaccharide. In some embodiments, the polysaccharide is present in the composition at about 0.3 M or about 60 mM. In some embodiments, any non-excipient component of the composition has a size of less than about 0.2 microns. In some embodiments, the composition is sterile by USP <71>. In some embodiments, the composition is endotoxin USP <85> free. In some embodiments, the composition is negative for mycoplasma DNA. In some embodiments, the composition is cell-free. In some embodiments, the composition is stored between -80 °C and -60 °C. In some embodiments, theAttorney Docket No.66309-727.601 composition is administered within 6 hours of thaw when maintained at ambient temperature. In some embodiments, the composition is present in a glass vial. In some embodiments, the composition is formulated for intravenous administration. In some embodiments, the composition has a pH of about 6 to about 7.5. In some embodiments, the BM-MSCs are negative for CD14, CD31, CD34, and CD45. In some embodiments, the BM-MSCs are positive for CD73, CD105, CD166, and CD90. In some embodiments, the BM-MSCs are capable of undergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, the BM-MSCs are obtained from an iliac crest aspiration of a single donor. In some embodiments, a concentration of the one or more EVs in the composition is measured by nanoparticle tracking analysis (NTA). In some embodiments, the NTA comprises light scatter and fluorescence evaluation. In some embodiments, the concentration of the one or more EVs is about 10 billion to about 250 billion EVs per mL of the composition. In some embodiments, the concentration of the one or more EVs is at about 1 billion to about 40 billion EVs per ml of the composition. In some embodiments, the one or more EVs have an average diameter of about 30 nm to about 170 nm. In some embodiments, a total protein concentration of the composition is about 10 to about 40 µg per ml of the composition. In some embodiments, a total protein concentration of the composition is about 1.5 to about 6 µg per ml of the composition. In some embodiments, the total protein concentration is measured by ELISA. In some embodiments, the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, and Let-7. In some embodiments, the one or more EVs comprise TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), and MIF (Macrophage migration inhibitory factor).

[0009] Disclosed herein are methods and compositions for increasing a level of anti-Mullerianhormone in a subject. In some embodiments, a method disclosed herein comprises administering to the subject a composition comprising TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocyte-macrophage colony-stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2),Attorney Docket No.66309-727.601 IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein- 3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein- glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), Transferrin, or a combination of two or more thereof. In some embodiments, the subject has a history of infertility. In some embodiments, the subject is from 21 to 50 years of age. In some embodiments, the subject is female. In some embodiments, the subject has a history of one or more failed in vitro fertilization (IVF) cycles. In some embodiments, the subject has a lower level of anti-Mullerian hormone as compared to a subject without a history of infertility. In some embodiments, the subject has premature ovarian failure (POF). In some embodiments, the subject has polycystic ovary syndrome (PCOS). In some embodiments, the subject has one or more of the following symptoms selected from the group consisting of: endometriosis, a thyroid disorder, a blocked fallopian tube, a damaged fallopian tube, pelvic inflammatory disease (PID), a high follicle-stimulating hormone (FSH) level, a low estrogen level, a fibroid, a polyp, an autoimmune disorder, a congenital uterine abnormality, a sexually transmitted infection (STI), a hormonal imbalance, a prolactin imbalance, a cortisol imbalance, a gonadotropin-releasing hormone (GnRH) deficiency, inability to conceive, irregular menstrual periods, absent menstrual periods, painful menstrual periods, cramping, heavy or prolonged menstrual bleeding, endometriosis, hormonal fluctuations, mood disturbances, sudden fluctuations in weight, sleep disturbances, and fatigue, or any combination thereof. In some embodiments, the subject is not menopausal or post-menopausal. In some embodiments, the administering comprises intravenous administration. In some embodiments, the intravenous administration is carried out over the course of 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments, the composition is prepared by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM- MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, a method disclosed herein comprises preparing the composition prior to the administering, wherein the preparing the composition occurs by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media havingAttorney Docket No.66309-727.601 a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition. In some embodiments, wherein the culture media is serum-free. In some embodiments, the culture media has a glucose concentration below 4.5 g / L. In some embodiments, formulating the MSC conditioned media comprises exchanging the conditioned media for a pharmaceutically acceptable formulation. In some embodiments, the pharmaceutically acceptable formulation comprises saline. In some embodiments, the composition comprises at least 6x1010to 8x1010extracellular vesicles per mL and is administered at a dose of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml. In some embodiments, the dose is administered in combination with normal saline at a final volume of 100 mL. In some embodiments, the composition comprises saline (0.9% sodium chloride). In some embodiments, the saline is present in the composition at about 80% to about 95% saline. In some embodiments, the composition comprises sodium chloride, sodium lactate, potassium chloride, and calcium chloride. In some embodiments, the molecular weight of any non- excipient component of the composition is greater than about 10 kDa (kilodaltons). In some embodiments, the composition comprises a polysaccharide. In some embodiments, the polysaccharide is present in the composition at about 0.3 M or about 60 mM. In some embodiments, any non-excipient component of the composition has a size of less than about 0.2 microns. In some embodiments, the composition is sterile by USP <71>. In some embodiments, the composition is endotoxin USP <85> free. In some embodiments, the composition is negative for mycoplasma DNA. In some embodiments, the composition is cell-free. In some embodiments, the composition is stored between -80 °C and -60 °C. In some embodiments, the composition is administered within 6 hours of thaw when maintained at ambient temperature. In some embodiments, the composition is present in a glass vial. In some embodiments, the composition is formulated for intravenous administration. In some embodiments, the composition has a pH of about 6 to about 7.5. In some embodiments, the BM-MSCs are negative for CD14, CD31, CD34, and CD45. In some embodiments, the BM-MSCs are positive for CD73, CD105, CD166, and CD90. In some embodiments, the BM-MSCs are capable of undergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, the BM-MSCs are obtained from an iliac crest aspiration of a single donor. In some embodiments, a concentration of the one or more EVs in the composition is measured by nanoparticle tracking analysis (NTA). In some embodiments, the NTA comprises light scatter and fluorescence evaluation. In some embodiments, the concentration of the one or more EVs is about 10 billion to about 250 billion EVs per mL of the composition. In some embodiments, the concentration of the one or more EVs is at about 1 billion to about 40 billion EVs per ml of the composition. In some embodiments, the one orAttorney Docket No.66309-727.601 more EVs have an average diameter of about 30 nm to about 170 nm. In some embodiments, a total protein concentration of the composition is about 10 to about 40 µg per ml of the composition. In some embodiments, a total protein concentration of the composition is about 1.5 to about 6 µg per ml of the composition. In some embodiments, the total protein concentration ismeasured by ELISA. In some embodiments, the one or more EVs comprise hsa-miR-21-5p,miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, and Let-7. In some embodiments, the one or more EVs comprise TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), and MIF (Macrophage migration inhibitory factor). INCORPORATION BY REFERENCE

[0010] Each patent, publication, and non-patent literature cited in the application is herebyincorporated by reference in its entirety as if each was incorporated by reference individually. To the extent publications and patents or patent applications incorporated by reference contradict the disclosure contained in the specification, the specification is intended to supersede and / or take precedence over any such contradictory material. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The features of the present disclosure are set forth with particularity in the appendedclaims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which:

[0012] FIGS. 1A-B show serum anti-Mullerian hormone (AMH) levels for subjects 101 through110 at day 0 and day 60. FIG.1A shows individual subject serum AMH levels plotted to show group mean (dark bar) with 95% confidence intervals (light whiskers). FIG.1B shows individual serum AMH levels plotted as continuous lines to illustrate subject heterogeneity in absolute levels and in response to the therapeutic composition over 60 days.Attorney Docket No.66309-727.601 DETAILED DESCRIPTION

[0013] Provided herein are compositions and methods for treating and managing symptoms offemale infertility. I. Compositions

[0014] Example compositions herein comprise an extracellular vesicle (EV) and / or a protein.The EV may originate from a mesenchymal stem cell (MSC). The protein may originate from a MSC. In an exemplary embodiment, the MSC is a bone marrow MSC (BM-MSC). The EV and / or protein may be purified or otherwise separated from the MSC growth and / or culturing condition from which the EV and / or protein was secreted into. Purified may include partially purified, such that some of the MSC growth and / or culturing condition is present in the composition. The composition may be formulated into an aqueous solution for intravenous administration. The composition may be referred to as a therapeutic composition. A. Extracellular Vesicles

[0015] Extracellular vesicles (EV) are small membrane bound spheres containing proteins andRNA (of which exosomes are a subset). Exosomes are small lipid bilayer vesicles secreted by cells that lack a nucleus and cannot replicate. Other EV populations are derived directly from the plasma membrane or are formed during apoptosis (apoptotic bodies). Disclosed herein are compositions comprising an EV. In example embodiments, the EV is an exosome. Embodiments of an EV herein have a diameter of about 20 nm to about 200 nm. In some embodiments, the diameter is measured by nanoparticle tracking analysis (NTA).

[0016] The number of EVs within a composition may be about 10 billion to about 250 billionEVs per mL when suspended. The suspension may be diluted for intravenous administration, wherein the EVs within the composition may be about 1 billion to about 40 billion EVs per mL.

[0017] In some embodiments, the EV has a phenotype of CD63+ CD9– and CD81–. In someembodiments, at least 70, 75, 80, 85, 90, 91, 92, 93, 94, or 95% of the EVs are CD63+CD9–and CD81–. In some embodiments, at least 50, 60, 70, 80, 85, 90, 91, 92, 93, 94, or 95% of the EVs are CD9–. In some embodiments, at least 50, 60, 70, 80, 85, 90, 91, 92, 93, 94, or 95% of the EVs are CD81–.

[0018] In some embodiments, the EV is produced from a MSC. The MSC may be a bonemarrow MSC. The MSC may be a human MSC. In some embodiments, the EV is produced from a MSC that has the capacity to undergo trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes. In some embodiments, the MSC is positive for CD73, CD105, CD166, and CD90 and is negative for CD14, CD31, CD34, and CD45.Attorney Docket No.66309-727.601

[0019] In some embodiments, EVs are analyzed via light scatter and fluorescence evaluation(e.g., NanoSight, Malvern Panalytical Ltd., United Kingdom). In some embodiments, the EVs are characterized by single particle interferometric reflectance imaging sensor technology to visualize and quantify fluorescent antibody-labeled particles (e.g., NanoView Biosciences, Boston, MA).

[0020] In some embodiments, the EV comprises a peptide or protein. In some embodiments, theEV comprises a nucleic acid. Nucleic acids include ribonucleic acids (RNA), such as siRNA, shRNA, and microRNA (miRNA).

[0021] In some embodiments, the EV comprises one or more proteins (e.g., 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100). Non-limiting examples of the one or more proteins include: bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD63 antigen, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Ferritin, Furin, GM-CSF Ra (Granulocyte-macrophage colony- stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein-3), IGFBP-4 (Insulin-like growth factor binding protein- 4), IGFBP-6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, MIF (Macrophage migration inhibitory factor), OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein-glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), Transferrin, and the proteins of Table 1. An EV that comprises the one or more of the proteins may include the one or more proteins within the EV. An EV that comprises one or more of the proteins may include the one or more proteins anchored within the EV. An EV that comprises the one or more of the proteins may be associated with the outside of the EV.

[0022] Table 1 below provides additional information regarding non-limiting examples ofproteins that may be comprised within an EV or EV composition of the present disclosure.Attorney Docket No.66309-727.601 Table 1 Protein Names , s ) , n, h n - 1 2; , 8 ,5, pericentrin-1Attorney Docket No.66309-727.601 Protein Names , s ) d e- 2, e , - - 8 e- - A , , n y - h dinduced , , , a CSD2, CSD3, ig-h3, kerato-Attorney Docket No.66309-727.601 Protein Names , s ) - - 4, h - g 1 ) 0 3 - - , - hn 6, IGFBP-6Attorney Docket No.66309-727.601 Protein Names , s ) , - - - ; A - - 1, , dr protein Bam32, B-Attorney Docket No.66309-727.601 Protein Names , s ) a io h n h h e a- - e ,a receptor, platelet-Attorney Docket No.66309-727.601 Protein Names , s ) T n - c , - - ,chain, GMCSFR-Attorney Docket No.66309-727.601 Protein Names , s ) h n - - y, EC:5.3.2.1Attorney Docket No.66309-727.601 Protein Names , s ) e 4 , y or 1 h n 4A), EC:3.4.24.79Attorney Docket No.66309-727.601 Protein Names , s ) - at 1 - A ll n e e c e - V , e, thioredoxin-Attorney Docket No.66309-727.601 Protein Names , s ) er 3 d r ; - , n r2 (TIMP-2)Attorney Docket No.66309-727.601 Protein Names , s ) , - or r - se l n e e , e e eENPP-2 ENPP2 Q13822 , , , pyrophosphatase / p AUTOTAXIN, ectonucleotideAttorney Docket No.66309-727.601 Protein Names , s ) h D - 5, t t , s , n lt eisolog, multipleAttorney Docket No.66309-727.601 Protein Names , s ) - fa - C , e I e a-, a, receptor subunit α IL-20RA, cytokineAttorney Docket No.66309-727.601 Protein Names , s ) e i , - n t , , r - 2 e r Ein , alfa, epoetin alpha-Attorney Docket No.66309-727.601 Protein Names , s ) r ll r rFABP6 FABP6 P51161 , , , , protein 6 BALB, I-BAP, fatty acid-bindingAttorney Docket No.66309-727.601 Protein Names , s ) - 5 ), - - 0 e c 2 4, r , g s ), dArtemin ARTN Q5T4W7 artemin , , , , EVN, NBN neublastinAttorney Docket No.66309-727.601 Protein Names , s ) e , e g 2 n e y t- oralpha-inducedAttorney Docket No.66309-727.601 Protein Names , s ) g A g g 9, n ), er rg ligand receptor 1Attorney Docket No.66309-727.601 Protein Names , s ) ; 1 2 d - d - c d - h n t 1 tr factor 7 SOX-7Attorney Docket No.66309-727.601 Protein Names , s ) y c - y e y s l- 2 , n , ), nn C), granulin-6Attorney Docket No.66309-727.601 Protein Names , s ) n ), a , - - a 1 n k e -2 (FEEL-2),Attorney Docket No.66309-727.601 Protein Names , s ) r 0 2 r , n- 2, - - , e d d(FKBP-5), FKBP54Attorney Docket No.66309-727.601 Protein Names , s ) - 4, , 1 4 g , - g L, g c lreceptor C , , Unc-5 homolog C unc-5 homolog 3,Attorney Docket No.66309-727.601 Protein Names , s ) t e r l - n, r I , I 6 2, SEMA6A1 (SEMA6A-1)Attorney Docket No.66309-727.601 Protein Names , s ) h - , 8 , / e - - e n e ll e(dipeptidylAttorney Docket No.66309-727.601 Protein Names , s ) e le r r - 2 n C, l 3 ia 0 e, CARD-containingAttorney Docket No.66309-727.601 Protein Names , s ) d - .- er s , , er , A A, pregnancy-Attorney Docket No.66309-727.601 Protein Names , s ) n n - F- l h m - dbeta A , , A inhibin, inhibin A, chain, erythroidAttorney Docket No.66309-727.601 Protein Names , s ) g , n r 2 n n , ; l - see lymphocyteAttorney Docket No.66309-727.601 Protein Names , s ) s s s s x 1 a - eproteinAttorney Docket No.66309-727.601 Protein Names , s ) r R - l - t e - r , - r incell antigen antigen CD34,Attorney Docket No.66309-727.601 Protein Names , s ) h- ta 2, r e d , ,n, macrophage-Attorney Docket No.66309-727.601 Protein Names , s ) , , , , , e , r r - er - 2 er eCD30, CD30Attorney Docket No.66309-727.601 Protein Names , s ) e 1 e n- e l e d i n er 4 ),Nidogen-1 NID1 P14543 nidogen 1 NID, ENTACTIN , , entactinAttorney Docket No.66309-727.601 Protein Names , s ) r, li d e- 1, e , er , - llg lectin 11, Siglec-11Attorney Docket No.66309-727.601 Protein Names , s ) - n 0 d , II A , - - y - ,domain 2 , , g beta, PH30-β, protein 2, ADAM 2,Attorney Docket No.66309-727.601 Protein Names , s ) n 0, 7, - n 8, n 9, - si h a , ,metalloprotease, αAttorney Docket No.66309-727.601 Protein Names , s ) e - e - , e - eIII)Attorney Docket No.66309-727.601 Protein Names , s ) d c - - e - e )- like disintegrin-likeAttorney Docket No.66309-727.601 Protein Names , s ) - e ) ) n - - r R B T, 2 ndin-2Attorney Docket No.66309-727.601 Protein Names , s ) , a- r- - - , , id , , , -Granzyme B GZMB P10144 granzyme B , , , , 1, CGL1, CSP-B, cathepsin G-like 1Attorney Docket No.66309-727.601 Protein Names , s ) n n e a d d , n , , 0 e e - - - ,sulfotransferase 2Attorney Docket No.66309-727.601 Protein Names , s ) 2 - n , n - e e - d- e -, n alpha chain, LH andAttorney Docket No.66309-727.601 Protein Names , s ) a , e , s d 2 r r , , - , c t -, , beta, IFN β interferon β 1,Attorney Docket No.66309-727.601 Protein Names , s ) n r r 1, - al - 4 , r, , g rreceptor 2 , SorCS2104 kDaAttorney Docket No.66309-727.601 Protein Names , s ) e er e - l r - , icg protein 8Attorney Docket No.66309-727.601 Protein Names , s ) er d - - r - 0 - e eLangerin CD207 Q9UJ71 CD207 molecule , Langerin domain family 4Attorney Docket No.66309-727.601 Protein Names , s ) n, r α- e - , - - , ), eD SEMA4D Q928 , , , 8, 4 54 semaphorin 4D C9orf164, CD100, BB18, GR3, CD100Attorney Docket No.66309-727.601 Protein Names , s ) g m 1, - er - d d , , 2, d , xTCSF, BASIGIN, inducerAttorney Docket No.66309-727.601 Protein Names , s ) d , n n, n e ), R α e ), - c n- y - e yy ltransferase 2Attorney Docket No.66309-727.601 Protein Names , s ) l ic l - e T r a r d r r , , 3,p53Attorney Docket No.66309-727.601 Protein Names , s ) e , 2 n n, n p 1 e F , , e e , .- n, SIR2-like protein 1Attorney Docket No.66309-727.601 Protein Names , s ) e e r e 5) , e , , t- r - ,(VAP-33)Attorney Docket No.66309-727.601 Protein Names , s ) , / e e - - 3 e .-... , EC:1.1.1.62Attorney Docket No.66309-727.601 Protein Names , s ) n n - T r , d , aBLAST1, MEM- antigen, B-Attorney Docket No.66309-727.601 Protein Names , s ) e , er or f r er, ... , methylumbelliferyl-Attorney Docket No.66309-727.601 Protein Names , s ) e - d r n, , , 2, n- - - , - , ,e, IMP-specific 5'-Attorney Docket No.66309-727.601 Protein Names , s ) , 5 - , ic n n e, 1B , , P28-ICK cyclin-dependentAttorney Docket No.66309-727.601 Protein Names , s ) 7, - er r, r , V, , - , ar m), HACBP, grp60Attorney Docket No.66309-727.601 Protein Names , s ) er g g g r a - 5, l , e , , e 1 6,surface associated LOC100293231, related tumorAttorney Docket No.66309-727.601 Protein Names , s ) - , e it a t 2, , g s ), d io , ) r -S52, processedAttorney Docket No.66309-727.601 Protein Names , s ) r y or e e e e d ) , - - n - -precursor, heparanAttorney Docket No.66309-727.601 Protein Names , s ) - 1 1 y e ) er - n s r h o nhomology to p53,Attorney Docket No.66309-727.601 Protein Names , s ) , n - d n , e , t ,, , EC:3.1.1.1),Attorney Docket No.66309-727.601 Protein Names , s ) l- e a , f - 1 , h n e, EC:3.4.21.108Attorney Docket No.66309-727.601 Protein Names , s ) ss - - ) n 2, e - ,PSPD D, PSP-D, SP-D,Attorney Docket No.66309-727.601 Protein Names , s ) D , a d d r , ar n l - eFLT4Attorney Docket No.66309-727.601 Protein Names , s ) , ), h h , g 2 er , er , er n r e ), 3, e E3 ubiquitinAttorney Docket No.66309-727.601 Protein Names , s ) of , A , - , - r L 5 - , ereceptor), MIS typeAttorney Docket No.66309-727.601 Protein Names , s ) I, or ), e e 3 - - e - e a c l - - n, , interleukin-1Attorney Docket No.66309-727.601 Protein Names , s ) 3, 6 t 3, - e , c s e e , e g ut , ,, receptor 3)Attorney Docket No.66309-727.601 Protein Names , s ) l F, r e 3 - r, f , a -, , urokinase R,Attorney Docket No.66309-727.601 Protein Names , s ) rIR , n 6, l ar l, , carcinoembryoniccAttorney Docket No.66309-727.601 Protein Names , s ) d 1 l- - e n , , 1 2activation CDw150 activation molecule,Attorney Docket No.66309-727.601 Protein Names , s ) e - n ), e , e, e antigen MAX.3,Attorney Docket No.66309-727.601 Protein Names , s ) e - - c er , l ),, , , PRO2222 protein (PRBP,Attorney Docket No.66309-727.601 Protein Names , s ) l- l- l- r y ), e n - , - - - 1 r r , e rN-Cadherin CDH2 P19022 cadherin 2 , , , l ARVD14, CD325, cadherin, N-Attorney Docket No.66309-727.601 Protein Names , s ) , - g - g n - - - 1 , - - -AMSH STAMBP O95630 , , protein LOC100507148 protein, associatedAttorney Docket No.66309-727.601 Protein Names , s ) - n n ε I, 2, F - ,, n CD28, CD28Attorney Docket No.66309-727.601 Protein Names , s ) , e - - - , rt u y- β - - E15 serin , , e ase e protease 15 protease 7,Attorney Docket No.66309-727.601 Protein Names , s ) , n- n, , I, io e - l lFLJ31125, β ark2 , EC:2.7.11.15Attorney Docket No.66309-727.601 Protein Names , s ) G - n e - e ) - - g ), eGalectin-2 LGALS2 P05162 galectin 2 HL14, GAL2 , , beta-galactoside-Attorney Docket No.66309-727.601 Protein Names , s ) - - - , e .- n , , , 2 a, 2IL-33 IL33 O95760 interleukin 33 , , , - IL1F11, NF-HEV, 33, interleukin-1Attorney Docket No.66309-727.601 Protein Names , s ) r s - 3 - 1 e c 1 e 5, r , r , n , L - e, SCM-1Attorney Docket No.66309-727.601 Protein Names , s ) y n , ), m - - - 4 n , 2 ,, cytokine receptor-Attorney Docket No.66309-727.601 Protein Names , s ) l er g , it , L- 4, ), nprotein RACK17Attorney Docket No.66309-727.601 Protein Names , s ) n e , ll , n - - , r ), - , r R, y IL-2 receptorAttorney Docket No.66309-727.601 Protein Names , s ) , F T - c s n c D 4 g r 1 e u-, endothelial cellAttorney Docket No.66309-727.601 Protein Names , s ) 1 h c n - , - , , g ), - , - 1 er, , or TRANCE-R, receptor (ODFR),Attorney Docket No.66309-727.601 Protein Names , s ) f , / e - e ), e - - e c 3 e 3, r , ,amidohydrolase 2,Attorney Docket No.66309-727.601 Protein Names , s ) l e C, C n n , I , , - n e - d αprecursorAttorney Docket No.66309-727.601 Protein Names , s ) r - ), r 7 , , , a , 1, e e -POGLUT1Attorney Docket No.66309-727.601 Protein Names , s ) t - , n 6 in n d 3 3, f 1, e er ,, , , , urokinase u-PA activator, u-Attorney Docket No.66309-727.601 Protein Names , s ) n n al , t e e ), r , , H l, , , - 1 PECAM-1, 1, EndoCAM,Attorney Docket No.66309-727.601 Protein Names , s ) 6 f- , er , n n - e e - d- e -hormone alpha,Attorney Docket No.66309-727.601 Protein Names , s ) , , H α t e - e x 2 , , - ,, ng protein (L36LBP),Attorney Docket No.66309-727.601 Protein Names , s ) or d - ll t T 3 r e, , ,n, hepatocyte growthAttorney Docket No.66309-727.601 Protein Names , s ) 2, N , - t 1 - , , e - e - - d, developmentally-Attorney Docket No.66309-727.601 Protein Names , s ) n or s, n n524Attorney Docket No.66309-727.601 Protein Names , s ) - 2, g ), - a- , r, e- - r- - - ), er ,, TAXAttorney Docket No.66309-727.601 Protein Names , s ) n t r - 2, - - , r ll - d - n er r, - R4), TRAILAttorney Docket No.66309-727.601 Protein Names , s ) er 1 c n , s e e e- n 1,member 4Attorney Docket No.66309-727.601 Protein Names , s ) io in r ur , 2, d, neurotrophicAttorney Docket No.66309-727.601 Protein Names , s ) , d h 1 h - g 1 , 7 e 7 8KIAA1777, homolog DAttorney Docket No.66309-727.601 Protein Names , s ) , , te - - 1 r R t , 4 ,XEDAR EDA2R Q9HAV5 , receptor EDAA2R, factor receptorAttorney Docket No.66309-727.601 Protein Names , s ) er - r r r 1, or -- FABP, AFABP),Attorney Docket No.66309-727.601 Protein Names , s ) e er r 1 ir e r, r - - - eKIAA1867, , - like protein 2,Attorney Docket No.66309-727.601 Protein Names , s ) 3 t 5 e- n, r e - e 6 n B, n- e 2 er 6RTN4R , , r, Receptor Q9BZR6 receptor UNQ330 / PRO526 Nogo receptorAttorney Docket No.66309-727.601 Protein Names , s ) , l - e - al e- - 5 , 5 - - td 2 2 CASPR2, CDFE, protein-like 2, cellAttorney Docket No.66309-727.601 Protein Names , s ) y - d e- 1, e , , - e ,(GLP-1(7-36)),Attorney Docket No.66309-727.601 Protein Names , s ) e e , e e e n t r - e d - c - -protein 2 , , - N2DL2, alpha, NKG2DAttorney Docket No.66309-727.601 Protein Names , s ) , e e 1, r l, , EC:2.7.10.1Attorney Docket No.66309-727.601 Protein Names , s ) 0 , d 1 A , , n n n, l , e n 4, - ,, WFDC2 isoform 3Attorney Docket No.66309-727.601 Protein Names , s ) - c y - e y er d hform, CD14Attorney Docket No.66309-727.601 Protein Names , s ) ) - C T- t r - - , er r -, p75, CD328Attorney Docket No.66309-727.601 Protein Names , s ) r, 5 l er s - er , n d 2, n g - -, CXCL12 isoform 1Attorney Docket No.66309-727.601 Protein Names , s ) a , F t- - n er n , n -e sarcoma viralAttorney Docket No.66309-727.601 Protein Names , s ) , 1 T - - y e 1 1 l ), - y n -, endothelial cellAttorney Docket No.66309-727.601 Protein Names , s ) 2 er - g e, d g g , , l- - n, RANTES(4-68)Attorney Docket No.66309-727.601 Protein Names , s ) , 2, n n n n n , n - - , r ,KIAA0012 n (TIL), CD281Attorney Docket No.66309-727.601 Protein Names , s ) er n n al y a 0 - , K er ,, affinity nerveAttorney Docket No.66309-727.601 Protein Names , s ) , , er y , , g , L n n, - e , y F ,it alpha, LIF receptorAttorney Docket No.66309-727.601 Protein Names , s ) er ), - , e n R 6 H K ,e 4, CD244Attorney Docket No.66309-727.601 Protein Names , s ) , - α, y r n - y , nHNFJ, HNFJ1, , n secreted formAttorney Docket No.66309-727.601 Protein Names , s ) , d - - d 1 d 2, , n g -, - 1-alpha(3-67)Attorney Docket No.66309-727.601 Protein Names , s ) n - e n , - h h - g 3 dn LFA-3, CD58Attorney Docket No.66309-727.601 Protein Names , s ) 3 - n 0 F t n , e S t n , - er - ,protein N-productAttorney Docket No.66309-727.601 Protein Names , s ) - n , a - - gmyeloid cells 2 , , DSP-7 on monocytes 2Attorney Docket No.66309-727.601 Protein Names , s ) e 3 d, e , n ) - n e n, - e ), y 3 -, - alpha(6-73)Attorney Docket No.66309-727.601 Protein Names , s ) 2, s n, 2, n r - t C 5, n α, 1, -, ic acid receptorAttorney Docket No.66309-727.601 Protein Names , s ) n e d r g , e C , d 2, - ),, beta-Attorney Docket No.66309-727.601 Protein Names , s ) , , 2 - - r n er ss 1 al , - d , K, , e EC:3.7.1.3Attorney Docket No.66309-727.601 Protein Names , s ) i I r r r n, r n , , , - eprotein 4 (MIP-4),Attorney Docket No.66309-727.601 Protein Names , s ) d - ), n P a- r , - erTNFR4 , maturation protein,Attorney Docket No.66309-727.601 Protein Names , s ) , 1, ), or 6 - - 4,cytokine A16Attorney Docket No.66309-727.601 Protein Names , s ) h h a, , - e y t , er ,protein 1 C2Attorney Docket No.66309-727.601 Protein Names , s ) ol is - L ly - , - d r 4, e n - n erLOC100131742 (Herpesvirus entryAttorney Docket No.66309-727.601 Protein Names , s ) ), e h E- , - - , - ,e, AADC, DOPAAttorney Docket No.66309-727.601 Protein Names , s ) 8 n - e n , 3 3 C 3 , n G 0 F tpeptidase 3 , , , , KLK2A1 gamma-Attorney Docket No.66309-727.601 Protein Names , s ) - P er d , r - l-- activating proteinAttorney Docket No.66309-727.601 Protein Names , s ) , y a, n - 1, - g n n m - d , P- - n, MIP-1-beta(3-69)Attorney Docket No.66309-727.601 Protein Names , s ) r R , e - , n r 1 - , mI, IL-8(1-77),Attorney Docket No.66309-727.601 Protein Names , s ) , - I, I, d ) m m - a e e n- , 8,acetyl-beta-D-Attorney Docket No.66309-727.601 Protein Names , s ) , , , , - g n e , , 3 -superfamily memberAttorney Docket No.66309-727.601 Protein Names , s ) le 1, d - - - er , 1, , 3IL-13 IL13 P35225 interleukin 13 , , , - NC30 13Attorney Docket No.66309-727.601

[0023] In some embodiments, the EV comprises one or more nucleic acids (e.g., 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100). Non-limiting examples of the one or more nucleic acids include: hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-29a-3p, hsa-miR- 29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, and hsa-miR-940. miRNA sequences may be obtained from www.mirbase.org. An EV that comprises the one or more of the nucleic acids may include the one or more nucleic acids within the EV. B. Proteins

[0024] Disclosed herein are compositions comprising a protein. The protein may be independentof an EV. For instance, the protein may not be present within the EV or within the membrane of the EV. The composition may comprise one or more proteins (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100). The protein may be present as a monomer or multimer. The protein may have a molecular weight (as a monomer or multimer as applicable) of at least about 10 kDa (kilodalton). Non-limiting examples of the one or more proteins include: bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD63 antigen, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Ferritin, Furin, GM-CSF Ra (Granulocyte-macrophage colony-stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein-3), IGFBP-4 (Insulin-like growth factor binding protein-4), IGFBP- 6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, MIF (Macrophage migration inhibitory factor), OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM- C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein-glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1Attorney Docket No.66309-727.601 (Thrombospondin 1), TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), Transferrin, and the proteins of Table 1.

[0025] In some embodiments, the one or more proteins comprises TIMP1. In someembodiments, the TIMP1 is present at about 200 pg / mL to about 80 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the one or more proteins comprises OPN. In some embodiments, the OPN is present at about 200 pg / mL to about 80 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the one or more proteins comprises IGFBP4. In some embodiments, the IGFBP4 is present at about 200 pg / mL to about 80 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the one or more proteins comprises osteonectin. In some embodiments, the osteonectin is present at about 200 pg / mL to about 80 ng / mL of the composition or about 30 pg / mL to about 12 ng / mL. In some embodiments, the concentration of the one or more proteins is measured by ELISA.

[0026] In some embodiments, the total protein concentration of the one or more proteins isabout 10 to about 40 µg per ml of the composition. For example, the concentration in frozen product. In some embodiments, the total protein concentration of the one or more proteins is about 1.5 to about 6 µg per ml of the composition. For example, the concentration in an intravenous solution for administration.

[0027] In some embodiments, compositions described herein can safely target one or more(e.g., more than hundreds) different biomolecular interactions or signaling pathways. In some embodiments, compositions described herein can treat one or more injuries or diseases caused by multiple etiologies. In some embodiments, compositions described herein can be used for treating a disease or condition without identifying the pathogen underlying the disease or condition, thus offering an advantage for treating a disease or condition caused by an emerging or previously unknown pathogen. II. Therapeutic Compositions

[0028] Compositions described herein may be administered in vivo in a pharmaceuticallyacceptable carrier. A pharmaceutically acceptable carrier may biologically suitable, i.e., the material may be administered to a subject, along with the EV and / or protein, without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the therapeutic composition in which it is contained. As a non-limiting example, the pharmaceutically acceptable carrier may comprise sodium chloride.

[0029] Parenteral administration of the composition, if used, is generally characterized byinjection. The injection may be an intravenous injection. The injection may involveAttorney Docket No.66309-727.601 administration of the composition over a period of about 60 minutes. The composition for injection may comprise saline, e.g., 0.9% saline (NaCl). The 0.9% saline may be about 85% to about 99% or more of the composition by volume. For instance, the composition comprises about 85% of 0.9% saline and about 15% of an EV and / or protein component. The EV and / or protein component may be formulated in an isotonic infusion solution. The EV and / or protein component may comprise sodium chloride. The EV and / or protein component may comprise sodium chloride, sodium lactate, potassium chloride, and calcium chloride. The EV and / or protein component may comprise sodium chloride (NaCl) at 100-150 mEq / L, sodium lactate (C₃H₅NaO₃) at 25-35 mEq / L, potassium chloride (KCl) at 2-6 mEq / L, calcium chloride (CaCl₂) at 1-5 mEq / L, and chloride ions (Cl⁻) at 80-130 mEq / L. The EV and / or protein component may comprise 0.1 M to 0.8 M polysaccharide. Non-limiting example polysaccharides include, but are not limited to, glucose, aldose (D-allose, D-altrose, D-mannose, etc.), glucopyranose, pentahydroxyhexanal, alpha-D-glucopyranoside dihydrate, a-D-glucopyranosyl-D-glucose, a-D- glucopyranosyl-dihydrate, polymer of P-D-glycopyranosyl units, P-D-fructofuranosyl a-D- glucopyranoside (anhydrous / dihydrate), β-D-galactopyranosyl-D-glucose, a-D-glucopyranosyl- a-D-glucopyranoside (anhydrous / dihydrate), galactose, pentoses (ribose, xylose, lyxose), dextrose, dodecacarbon monodecahydrate, fructose, sucrose, lactose, maltose, trehalose, agarose, D-galactosy1-0-(1-4)-anhydro-L-galactosyl, cellulose, starch, polyhydric alcohol, polyalcohol, alditol, erythritol, glycitol, glycerol, xylitol, and sorbitol. In some embodiments, the composition has a pH of about 6 to about 7.5.

[0030] In some embodiments, the composition is sterile by USP <71>. In some embodiments,the composition is endotoxin USP <85> free. In some embodiments, the composition is negative for mycoplasma DNA. In some embodiments, the composition is cell-free. In some embodiments, the composition is stored between -80 °C and -60 °C. In some embodiments, the composition is administered within 6 hours of thaw when maintained at ambient temperature. In some embodiments, the composition is stored in a glass vial. A. Methods of Composition Production

[0031] Compositions herein comprising an EV and a protein may include components producedfrom a MSC. The EV of the composition may be produced from a MSC. The protein of the composition may be produced from a MSC. The EV and / or protein may be produced from a MSC cultured at one or more of the following conditions: about 0.1% to about 5% oxygen, reduced or no serum, pH of about 5-7.5, reduced glucose, increased temperature, or any of these elements in various combinations. The MSC may be cultured under the aforementioned one or more conditions after the cell achieves confluency. The MSC may be cultured under theAttorney Docket No.66309-727.601 aforementioned one or more conditions after the cell is cultured at 37°C and 5% CO2. The EV and / or protein of the composition may be obtained from a method comprising: (1) growing the MSCs to 50-80% confluency, then (2) culturing the MSCs at one or more of the following conditions: about 1% to about 5% oxygen, reduced or no serum, reduced glucose, or increased temperature. The growing at step (1) may be performed at about 37°C and about 5% CO2. Culturing the MSCs may comprise introducing the cells to a culture media. The culture media may comprise basal media. For instance, basal media comprising amino acids, vitamins, and inorganic salts, and optionally a carbon source such as glucose. The culture media may comprise an isotonic infusion solution. In a non-limiting example embodiment, the culture media comprises a basal media and an isotonic infusion solution. The culture media may comprise basal media and sodium chloride. The culture media may comprise basal media, sodium chloride, sodium lactate, potassium chloride, and calcium chloride. The culture media may comprise a basal media and an isotonic infusion solution. The culture media may comprise basal media and sodium chloride, without serum. The culture media may comprise basal media, sodium chloride, sodium lactate, potassium chloride, and calcium chloride, without serum. The culturing at step (2) may occur over a period of 1, 2, 3, 4, 5, 6, or 7 days, wherein the oxygen and pH may change over time. At any time during the culturing step, the pH may be about 6.5 to about 7. At any time during the culturing step, the oxygen may be at about 0.5%, 1%, 1.5%, or 2%.

[0032] The EV and / or protein produced in the second culturing step (2) may be purified (e.g.,partially or entirely), from the culture media of the second culturing step (2). The purification may comprise formulating the EV and / or the protein into an EV and / or protein component. The EV and / or protein component may comprise an isotonic infusion solution. The EV and / or protein component may comprise sodium chloride. The EV and / or protein component may comprise sodium chloride, sodium lactate, potassium chloride, and calcium chloride. The EV and / or protein component may comprise sodium chloride (NaCl) at 100-150 mEq / L, sodium lactate (C₃H₅NaO₃) at 25-35 mEq / L, potassium chloride (KCl) at 2-6 mEq / L, calcium chloride (CaCl₂) at 1-5 mEq / L, and chloride ions (Cl⁻) at 80-130 mEq / L. The EV and / or protein component may comprise a polysaccharide, e.g., 0.1 M to 0.8 M polysaccharide. Non-limiting example polysaccharides include, but are not limited to, glucose, aldose (D-allose, D-altrose, D- mannose, etc.), glucopyranose, pentahydroxyhexanal, alpha-D-glucopyranoside dihydrate, a-D- glucopyranosyl-D-glucose, a-D-glucopyranosyl-dihydrate, polymer of P-D-glycopyranosyl units, P-D-fructofuranosyl a-D-glucopyranoside (anhydrous / dihydrate), β-D-galactopyranosyl-D- glucose, a-D-glucopyranosyl-a-D-glucopyranoside (anhydrous / dihydrate), galactose, pentoses (ribose, xylose, lyxose), dextrose, dodecacarbon monodecahydrate, fructose, sucrose, lactose,Attorney Docket No.66309-727.601 maltose, trehalose, agarose, D-galactosy1-0-(1-4)-anhydro-L-galactosyl, cellulose, starch, polyhydric alcohol, polyalcohol, alditol, erythritol, glycitol, glycerol, xylitol, and sorbitol. The amount of polysaccharide in the EV and / or protein component may be about 0.2 M to about 0.6 M, or about 0.3 M. In some embodiments, the EV and / or protein component is frozen. The frozen material may be thawed and combined with saline (e.g., 0.9% saline or sodium chloride) to generate an IV formulation for IV administration. The amount of polysaccharide in the IV formulation may be about 40 mM to about 80 mM, or about 60 mM.

[0033] The EV and / or protein component may be filter-sterilized. The EV and / or proteincomponent may be filter sterilized after and / or during formulation from the culture media into the EV and / or protein component. The EV and / or protein may be concentrated. The concentration may occur during and / or after purification and / or exchange from the culture media into the EV and / or protein component. The EV and / or protein may be frozen, e.g., after and / or during purification. If frozen, the EV and / or protein may be formulated with a cryoprotectant prior to freezing. The cryoprotectant may comprises a polysaccharide, e.g., as described above.

[0034] In some embodiments, reduced glucose is less than a normal control (e.g., 4.5 g / L). Forexample, the glucose reduction may be about 5% to about 15%, from about 10% to about 20%, from about 15% to about 25%, from about 20% to about 30%, or from about 25% to about 35% of the glucose in a normal control. In some embodiments, the reduced glucose is present in the MSC culture media (e.g., at step (2) as noted above) at a concentration of about 0.1, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, or 4.0 g / L, or a range between any two of these values. In some embodiments, glucose is present at a concentration of less than or no more than 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, or 4.5 g / L.

[0035] In some embodiments, the MSC is cultured, e.g., at step (2) at about 0.1% to about 5%oxygen, for example, about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5% oxygen. In an example embodiment, the oxygen is about 0.5% to about 1.5%, or about 1%.

[0036] The pH at which the MSC is cultured, e.g., at step (2), can be from about 6.0 to about7.4, for example, from 6.5 to about 7, or about 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, or 7.4.

[0037] The temperature of the culture environment, e.g., at step (2), may be raised relative tophysiologic homeostasis temperature (e.g., 37°C). In one aspect, the temperature of the culture can be 35.0, 35.1, 35.2, 35.3, 36.4, 35.5, 35.6, 35.7, 35.8, 35.9, 36.0, 36.1, 36.2, 36.3, 36.4, 36.5, 36.6, 36.7, 36.8, 36.9, 37.0,.37.1, 37.2, 37.3, 37.4, 37.5, 37.6, 37.7, 37.8, 37.9, 38.0, 38.1, 38.2,Attorney Docket No.66309-727.601 38.3, 38.4, 38.5, 38.6, 38.7, 38.8, 38.9, 39.0, 39.1, 39.2, 39.3, 39.4, 39.5, 39.6, 39.7, 39.8, 39.9, or 40.0°C.

[0038] In some embodiments, compositions described herein can be lyophilized for packagingand storing. In some embodiments, compositions described herein can be stored at ambient or room temperature (e.g., between 60°F and 75°F or between 15°C and 24°C). In some embodiments, the stability of compositions described herein at ambient or room temperature can provide advantages for logistics and / or delivery.III. Methods of treating infertility

[0039] Provided herein in some embodiments are methods and compositions for treatinginfertility. In some embodiments, the methods provided herein may be used to improve or alleviate symptoms of infertility. In some embodiments, infertility may comprise human infertility. In some embodiments, infertility may comprise female infertility.

[0040] In some embodiments, methods and compositions disclosed herein may be used to treatinfertility or symptoms thereof in a human subject. In some embodiments, a human subject may be from 21 to 50 years of age. In some embodiments, a human subject may be 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 years of age. In some embodiments, a human subject may be female. In some embodiments, a human subject may have a history of infertility. In some embodiments, a human subject may have a history of one or more failed in vitro fertilization (IVF) cycles. In some embodiments, a human subject with a history of infertility may have a lower level of anti-Mullerian hormone as compared to a human subject without a history of infertility. In some embodiments, a human subject is not pregnant. In some embodiments, a human subject is not menopausal. In some embodiments, a human subject is not post-menopausal. In some embodiments, a human subject has one or more of: premature ovarian failure (POF), polycystic ovary syndrome (PCOS), endometriosis, a thyroid disorder, a blocked fallopian tube, a damaged fallopian tube, pelvic inflammatory disease (PID), a high follicle-stimulating hormone (FSH) level, a low estrogen level, a fibroid, a polyp, an autoimmune disorder, a congenital uterine abnormality, a sexually transmitted infection (STI), a hormonal imbalance, and / or one or more failed repetitive IVF cycles. In some embodiments, a hormonal imbalance may comprise a prolactin imbalance, a cortisol imbalance, and / or a gonadotropin-releasing hormone (GnRH) deficiency. In some embodiments, a human subject may not have cancer. In some embodiments, symptoms of infertility may comprise but are not limited to inability to conceive, irregular menstrual periods, absent menstrual periods, painful menstrual periods, cramping, heavy or prolonged menstrualAttorney Docket No.66309-727.601 bleeding, endometriosis, hormonal fluctuations, mood disturbances, sudden fluctuations in weight, sleep disturbances, and fatigue.

[0041] In some embodiments, the compositions disclosed herein may be used in a method oftreating infertility in a human subject. In some embodiments, the compositions disclosed herein may be administered to the human subject. In some embodiments, the administration may be intravenous. In some embodiments, the administration may comprise an intravenous infusion. In some embodiments, the administration may be carried out over the course of several minutes. In some embodiments, the administration may be carried out over the course of 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 minutes. In some embodiments, the administration may be carried out over the course of greater than 60 minutes. In some embodiments, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 milliliters (mL) of the compositions disclosed herein may be administered at a time. In some embodiments, a dose of a composition disclosed herein may be administered. In some embodiments, a dose of a composition disclosed herein may comprise 1, 2, 3, 4, or 5 milliliters. In some embodiments, a dose of a composition disclosed herein may be combined with normal saline. In some embodiments, a dose of a composition disclosed herein may be combined with normal saline for a final volume of 100 milliliters. In some embodiments, a dose of 5 milliliters of a composition disclosed herein may be combined with 95 milliliters of normal saline. In some embodiments, the compositions disclosed herein may be administered once. In some embodiments, the compositions disclosed herein may be administered more than once.

[0042] In some embodiments, the compositions disclosed herein may comprise one or more of:bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD63 antigen, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Ferritin, Furin, GM-CSF Ra (Granulocyte-macrophage colony-stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein-3), IGFBP-4 (Insulin-like growth factor binding protein-4), IGFBP-6 (Insulin- like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-associated membrane glycoprotein 2), Lumican, MIF (Macrophage migration inhibitory factor), OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor),Attorney Docket No.66309-727.601 Sortilin, TGM4 (Protein-glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), TIMP-1 (Tissue inhibitor of MMPs 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), and Transferrin, or any combination thereof. In some embodiments, the compositions disclosed herein may comprise one or more of: hsa-miR-125b-5p, hsa-miR-145- 5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-21-5p,hsa-miR-221-3p, hsa-miR-222-3p, hsa- miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-27b-3p, hsa-miR-29a- 3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa- miR-424-5p, and hsa-miR-940, or any combination thereof.

[0043] In some embodiments, a cell-equivalent dose of a composition disclosed herein maycomprise from 0.7 to 7.0 million cells per kilogram. In some embodiments, a cell-equivalent dose of a composition disclosed herein may comprise 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7.0 million cells per kilogram. In some embodiments, a dose of a composition disclosed herein provides 9x1011to 1.2x1012extracellular vesicles or 5x1011to 1.5x1012, 6x1011to 1.4x1012, 7x1011to 1.3x1012, 8x1011to 1.2x1012, or 8x1011to 1.3x1012extracellular vesicles. In some embodiments, a composition disclosed herein is administered at a dose that provides at least 5x1011, 6x1011, 7x1011, 8x1011, 9x1011, 1x1012, 1.1x1012, 1.2x1012, 1.3x1012, 1.4x1012, or 1.5x1012extracellular vesicles. In some embodiments, a composition disclosed herein comprises 6x1010to 8x1010extracellular vesicles per mL and is administered at a dose of 5 to 20 mL.

[0044] In some embodiments, a level of anti-Mullerian hormone may be measured in a humansubject prior to the administration. In some embodiments, a level of anti-Mullerian hormone may be measured in a human subject after the administration. In some embodiments, a level of anti-Mullerian hormone may be measured in a human subject 30 days after the administration. In some embodiments, a level of anti-Mullerian hormone may be measured in a human subject 60 days after the administration. In some embodiments, a level of anti-Mullerian hormone in a human subject may be measured in a blood sample from the human subject. In some embodiments, a level of anti-Mullerian hormone in a human subject may be measured in serum. In some embodiments, a human subject with a history of infertility may have an anti-Mullerian hormone level of 0.0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, or 2.0 nanograms / milliliter. In some embodiments, a subject may have an anti- Mullerian hormone level of 0.0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, or 2.0 nanograms / milliliter after the administration. In some embodiments, a human subject may have an increase in a level of anti-Mullerian hormone of 0.0, 0.1, 0.2, 0.3,Attorney Docket No.66309-727.601 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, or 2.0 nanograms / milliliter after the administration. In some embodiments, a human subject may have an increase in anti- Mullerian hormone level for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or 1 year or more after the administration.

[0045] In some embodiments, the methods and compositions disclosed herein may be used totreat or improve symptoms of infertility in a human subject. In some embodiments, a human subject may have an increased level of anti-Mullerian hormone after administration of a composition disclosed herein as compared to before the administration. In some embodiments, a human subject may have an increased level of anti-Mullerian hormone 30 days after administration of a composition disclosed herein. In some embodiments, a human subject may have an increased level of anti-Mullerian hormone 60 days after administration of a composition disclosed herein. In some embodiments, a human subject may have an improvement in fertility after administration of a composition disclosed herein as compared to before administration.IV. Definitions

[0046] As used in this specification and the appended claims, the singular forms “a,” “an,” and“the” include plural referents unless the content clearly dictates otherwise. It should also be noted that the term “or” is generally employed in its sense including “and / or” unless the content clearly dictates otherwise. The terms “and / or” and “any combination thereof” and their grammatical equivalents as used herein, can be used interchangeably. These terms can convey that any combination is specifically contemplated. Solely for illustrative purposes, the following phrases “A, B, and / or C” or “A, B, C, or any combination thereof” can mean “A individually; B individually; C individually; A and B; B and C; A and C; and A, B, and C.” The term “or” can be used conjunctively or disjunctively, unless the context specifically refers to a disjunctive use.

[0047] The term “about” or “approximately” can mean within an acceptable error range for theparticular value, which may depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” can mean within 1 or more than 1 standard deviation. Alternatively, “about” can mean a range of up to 20%, up to 10%, up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, within 5-fold, or within 2-fold, of a value. Where particular values are described in the application and claims, unless otherwise stated the term “about” meaning within an acceptable error range for the particular value should be assumed.Attorney Docket No.66309-727.601

[0048] As used in this specification and claim(s), the words “comprising” (and any form ofcomprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method or composition of the present disclosure, and vice versa. Furthermore, compositions of the present disclosure can be used to achieve methods of the present disclosure.

[0049] Reference in the specification to “some embodiments,” “an embodiment,” “oneembodiment” or “other embodiments” means that a particular feature, structure, or characteristic described in connection with the embodiments is included in at least some embodiments, but not necessarily all embodiments, of the present disclosures. To facilitate an understanding of the present disclosure, a number of terms and phrases are defined below.

[0050] Certain specific details of this description are set forth in order to provide a thoroughunderstanding of various embodiments. However, one skilled in the art will understand that the present disclosure may be practiced without these details. In other instances, well-known techniques or methods have not been shown or described in detail to avoid unnecessarily obscuring descriptions of the embodiments. Unless the context requires otherwise, throughout the specification and claims which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense, that is, as “including, but not limited to.” Further, headings provided herein are for convenience only and do not interpret the scope or meaning of the claimed disclosure.

[0051] Although methods and materials similar or equivalent to those described herein can beused in the practice or testing of the present disclosure, suitable methods, and materials are described below. V. Examples

[0052] The following examples are put forth so as to provide those of ordinary skill in the artwith a complete disclosure and description of how the compounds, compositions, articles, devices and / or methods claimed herein are made and evaluated and are intended to be purely exemplary and are not intended to limit the disclosure. Efforts have been made to ensure accuracy with respect to numbers (e.g., amounts, temperature, etc.), but some errors and deviations should be accounted for. These examples are provided for illustrative purposes only and not to limit the scope of the claims provided herein.Attorney Docket No.66309-727.601 A. Example 1 – Production of Therapeutic Composition

[0053] An MSC secretome therapeutic composition (investigational medicinal product or“IMP”) was made by the following method: human bone marrow-derived MSCs were cultured in culture vessels with growth media to expand the MSC population. Growth media was then removed, and the cells were washed with PBS. The MSCs were then cultured at a pH below 7.0, less than 5% oxygen (e.g., to a final concentration of about 1% oxygen), and in a culture media comprising basal media and sodium chloride. The culture media comprising the MSC secretome was formulated into an isotonic infusion solution, such as sodium chloride, and filter sterilized. The production process for the IMP was done under current Good Manufacturing Practices and Current Good Tissue Practices.

[0054] The IMP was manufactured from the banked hBM-MSCs of a single donor under CGMPconditions and according to FDA Master File protocols. Each lot of the IMP was characterized by proteomic and miRNA characterization. Additionally, the size and quantity of EVs and the presence of a specific surface marker expression profile were confirmed.

[0055] The IMP comprises extracellular vesicles (EVs) that are acellular and nonimmunogenic,containing no nucleus or deoxyribonucleic acid (DNA).

[0056] The tetraspanin profile of extracellular vesicles present in the IMP was determined, andit was found that greater than 95% of the extracellular vesicles present in the composition were CD63+CD9–CD81–. The EVs were measured via Nanoparticle tracking analysis (NTA), having a median diameter of about 100 nm. At least 10 billion EVs per mL were calculated using NTA with fluorescent staining of EV membranes.

[0057] Protein content of the IMP was determined, and the following proteins were found to bepresent. Proteins were detected using an antibody-based sandwich ELISAs or by Luminex or Nanoview / Unchained Labs-based methods. The total concentration of certain proteins in the IMP was measured via ELISA, with each protein having a concentration of about 200 pg / mL to about 80 ng / mL. For instance, TIMP1, OPN, IGFBP4, and osteonectin were characterized. Non- limiting examples of proteins present in the IMP are shown in Table 2 below. Table 2bIG-H3 Transforming growth factor-beta-induced protein ig-h3Attorney Docket No.66309-727.601 Protein ID Full NameCD99 CD99 antigenAttorney Docket No.66309-727.601 Protein ID Full Name

[0058] The nucleic acid content of the IMP was determined, and the following nucleic acidswere found to be present: hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa- miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-29a-3p, hsa-miR-29c- 3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, and hsa- miR-940. RNA were measured using semi-quantitative PCR. Total RNA was analyzed by UV detection using a ClarioStar Plus microplate reader and BMG Labtech LVis plate adapter for nucleic acid quantification. Using this method, the amount of each RNA may be less than the limit of detection, or about 50 ng / mL. B. Example 2 – Prospective Study Evaluating Safety and Efficacy ofTherapeutic Product in Infertile Women

[0059] Objective: This prospective non-randomized, open-label pilot study addressed the safetyof a human bone marrow-derived extracellular vesicle (hBM-MSC EV) therapeutic product of Example 1 above as a potential treatment for human infertility. Subjects: Ten female subjects under evaluation for infertility at a single clinic location. Inclusion criteria were women aged 21 to 50 years, without medical comorbidities, with a history of infertility and / or previous failure of in-vitro fertilization attempts. Intervention: All subjects received a single intravenous administration of 5 mL of the therapeutic product. Main Outcome Measures: Outcome measures included observation or reporting of subject adverse events and laboratory measurement of serum anti-Mullerian hormone (AMH) levels. Baseline AMH levels were measured prior to infusion and again at 30- and 60-days post infusion. Results: No adverse events were detected during an observation period following infusion of the IP and none were reported during a subsequent 48-hour notification period. AMH levels and changes over 60 days were heterogeneous. Six of 10 subjects exhibited increased levels of AMH at 60 days.Attorney Docket No.66309-727.601 Conclusions: The therapeutic product was safe in this study and was able to elevate AMH levels in some patients, indicating its ability to reduce infertility in women.

[0060] Subject cohort: Ten subjects were enrolled in the study who met the following inclusioncriteria: adult females between ages 36 and 49 years with a history of infertility and / or previous IVF failures, no underlying co-morbidity factors, no history of severe allergic reactions, and subjects were afebrile and normotensive. Voluntary signature of informed consent by the subject or healthcare proxy was obtained. Any potential enrollee was excluded if they met any of the following exclusion criteria: subject was pregnant, positive history of any malignancy in previous five years, positive results in cancer screening panel, comprehensive metabolic panel abnormalities, or greater than 50 years of age or post-menopausal.

[0061] Subject Evaluation: The Evaluation Procedure was initiated with an intake consultationobtained remotely via Zoom, Skype, or phone contact. Preliminary history was obtained and labs ordered. Once lab results were obtained and reviewed by the investigator to rule out abnormality, a subject visit for dose administration was scheduled. The treated subject population presented with infertility diagnoses of premature ovarian failure (POF), polycystic ovary syndrome (PCOS), and repetitive failed IVF cycles.

[0062] Assessments: Subject testing performed prior to infusion included CBC, CMP, CancerScreening Panel and Anti-Mullerian Hormone (AMH) less than 10 days prior to infusion. Vitals were obtained on the day of infusion and an IV started by a Registered Nurse (RN). The infusion occurred over a period of 60 minutes. Vitals were then obtained post infusion and prior to discharge. Subjects were monitored for any adverse response for 30-60 minutes post infusion and given instructions to call with any physical concerns over the next 24-48 hours. Clinical laboratory testing was performed at 30- and 60-days post infusion to measure serum AMH levels.

[0063] Intervention: The dosing protocol comprised a single dose of the Example 1 therapeuticproduct. The single dose was 5 mL of the therapeutic product in 95 mL Normal Saline (NS) administered via an IV catheter per infusion. The product was administered in a 100 mL bag of NS over 30 to 60 minutes. The delivered 5 mL EV dose in a final volume of 100 mL NS is predicted to fall within the cell-equivalent dose range of 0.7 to 7 million cells / kg >95% of the time.

[0064] A manufacturing lot certificate of analysis is provided with each vial of the therapeuticproduct and a single lot of the product was utilized during the study. The product is documented to provide biological mediators that reduce inflammation, facilitate paracrine signaling and advance healing processes. Consistent with mechanism of action studies performed to assess the efficacy of MSCs, the secreted biomolecule milieu captured in the therapeutic product isAttorney Docket No.66309-727.601 complex, and the primary mode of action of such a pleiotropic therapeutic may be influenced by the environment into which it is introduced in the body.

[0065] Results: Subject characteristics: CMC and CBC lab results for all 10 treated subjectswere within normal and acceptable ranges prior to the treatment indicating that all subjects were healthy except for their documented infertility issues. No adverse events were detected during the infusion and recovery periods, and no adverse events were reported by the subjects over the 48-hour period following the infusion (Table 2).

[0066] Serum AMH levels: AMH level results for the 10 subjects are shown in FIGS. 1A-1B.Among subjects 101, 102, 103, 105, 107 and 108 an increase in AMH level was apparent 60 days after treatment as compared to the individual subject’s baseline level (day 0). The results suggest that the therapeutic product may have the potential to improve AMH levels and ovarian insufficiency in a subset of infertile subjects.

[0067] No adverse events associated with the therapeutic product were detected by theinvestigators during the intervention, and none were reported by the participants during the follow up period. These results are consistent with the acceptable safety profile reported for a higher dose of the therapeutic product administered in the hospital setting to severely ill acute COVID-19 subjects. C. Example 3 – (Prophetic) Treatment of Infertility and Associated Symptomsin Subjects Treated with the Therapeutic Product

[0068] Provided in this Example is the use of the composition disclosed herein as preparedaccording to Example 1 in the treatment of infertility in human subjects. The therapeutic product is administered via intravenous infusion to subjects with a history of infertility. Subjects between the ages of 21 and 50 are selected for treatment. Several subjects have a history of multiple failed IVF cycles and inability to conceive. Prior to administration, subjects are found to have relatively low levels of anti-Mullerian hormone and estrogen and a relatively high level of follicle-stimulating hormone relative to subjects who do not have a history of infertility. After administration, subjects are found to have increased levels of anti-Mullerian hormone and estrogen and a reduced level of follicle-stimulating hormone compared to before administration. Subjects report a reduction in symptoms including cramping, mood disturbance, and irregular menstrual cycle. Several subjects become pregnant within six months of administration of the therapeutic product.

[0069] The examples and embodiments described herein are for illustrative purposes only andvarious modifications or changes suggested to persons skilled in the art are to be included within the spirit and purview of this application.

Claims

Attorney Docket No.66309-727.601 CLAIMS WHAT IS CLAIMED IS:

1. A method of treating infertility or symptoms thereof in a subject, the method comprising:administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR- 27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa- miR-221-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa-miR-27a-3p, hsa-miR-29a- 3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa- miR-424-5p, or hsa-miR-940, or a combination of two or more thereof.

2. A method of treating infertility or symptoms thereof in a subject, the method comprising:administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein at least 80% of the EVs are CD63+, CD9-, CD81-.

3. A method of treating infertility or symptoms thereof in a subject, the method comprising:administering to the subject a composition comprising: TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocyte-macrophage colony-stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein-3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome- associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein-glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), Transferrin, or a combination of two or more thereof.

4. The method of any one of claims 1-3, wherein the subject has a history of infertility.

5. The method of any one of claims 1-4, wherein the subject is from 21 to 50 years of age.Attorney Docket No.66309-727.6016. The method of any one of claims 1-5, wherein the subject is female.

7. The method of any one of claims 1-6, wherein the subject has a history of one or morefailed in vitro fertilization (IVF) cycles.

8. The method of claim 4, wherein the subject has a lower level of anti-Mullerian hormoneas compared to a subject without a history of infertility.

9. The method of any one of claims 1-8, wherein the subject has premature ovarian failure(POF).

10. The method of any one of claims 1-9, wherein the subject has polycystic ovary syndrome(PCOS).

11. The method of any one of claims 1-10, wherein the subject has one or more of thefollowing symptoms selected from the group consisting of: endometriosis, a thyroid disorder, a blocked fallopian tube, a damaged fallopian tube, pelvic inflammatory disease (PID), a high follicle-stimulating hormone (FSH) level, a low estrogen level, a fibroid, a polyp, an autoimmune disorder, a congenital uterine abnormality, a sexually transmitted infection (STI), a hormonal imbalance, a prolactin imbalance, a cortisol imbalance, a gonadotropin-releasing hormone (GnRH) deficiency, inability to conceive, irregular menstrual periods, absent menstrual periods, painful menstrual periods, cramping, heavy or prolonged menstrual bleeding, endometriosis, hormonal fluctuations, mood disturbances, sudden fluctuations in weight, sleep disturbances, and fatigue, or any combination thereof.

12. The method of any one of claims 1-11, wherein the subject is not menopausal or post-menopausal.

13. The method of any one of claims 2-12, wherein the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR-145- 5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa- miR-23b-3p, hsa-miR-27a-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR- 320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, or hsa-miR-940, or a combination of two or more thereof.

14. The method of claim 1 or claim 2, wherein the composition comprises TIMP-1 (tissueinhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocyte-macrophage colony-stimulating factor receptorAttorney Docket No.66309-727.601 subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein-3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome- associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein-glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), Transferrin, or a combination of two or more thereof.

15. A method of increasing a level of anti-Mullerian hormone in a subject, the methodcomprising: administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein at least 80% of the EVs are CD63+, CD9-, CD81-.

16. A method of increasing a level of anti-Mullerian hormone in a subject, the methodcomprising: administering to the subject a composition comprising one or more extracellular vesicles (EVs), wherein the one or more EVs comprise comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, Let-7, hsa-miR-125b-5p, hsa-miR-145-5p, hsa-miR-191-5p, hsa-miR-199a-3p, hsa-miR-221-3p, hsa-miR-22-3p, hsa-miR-23a-3p, hsa-miR-23b-3p, hsa- miR-27a-3p, hsa-miR-29a-3p, hsa-miR-29c-3p, hsa-miR-31-5p, hsa-miR-320a, hsa-miR-34a-5p, hsa-miR-423-3p, hsa-miR-424-5p, or hsa-miR-940, or a combination of two or more thereof.

17. A method of increasing a level of anti-Mullerian hormone in a subject, the methodcomprising: administering to the subject a composition comprising TIMP-1 (tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), MIF (Macrophage migration inhibitory factor), bIG-H3 (Transforming growth factor-beta-induced protein ig-h3), CA2 (Carbonic anhydrase 2), Cathepsin B, CD81 antigen, CD9 antigen, CD99 antigen, CD109 antigen, CNTF (Ciliary neurotrophic factor), Decorin, DcR3 (Tumor necrosis factor receptor superfamily member 6B), Dkk-3 (Dickkopf-related protein 3), Endoglycan (podocalyxin-like protein 2), Furin, GM-CSF Ra (Granulocyte-macrophage colony-stimulating factor receptor subunit alpha), GPR115 (Adhesion G protein-coupled receptor F4), GP73 (Golgi membrane protein 1), HS3ST4 (Heparan sulfate glucosamine 3-O-sulfotransferase 4), IGF-2 (Insulin-like growth factor-2), IGFBP-2 (Insulin-like growth factor binding protein-2), IGFBP-3 (Insulin-like binding protein-3), IGFBP-6 (Insulin-like growth factor binding protein-6), IL-1 R6 (Interleukin 1 Receptor 6), ILFBP-4 (Insulin-like growth factor binding protein-4), LAMP2 (Lysosome-Attorney Docket No.66309-727.601 associated membrane glycoprotein 2), Lumican, OPN (Osteopontin), PAI-1 (Plasminogen activator inhibitor-1 or SERPINE 1), PCK1 (Phosphoenolpyruvate carboxykinase, cytosolic), PF4 (Platelet Factor 4), Periostin, PRDX4 (Peroxidredoxin-4), RGM-C (Hemojuvelin), Semaphorin 6C, Serpin B6, Serpin F1 (Pigment epithelium-derived factor), Sortilin, TGM4 (Protein-glutamine gamma-glutamyltransferase 4), Thrombomodulin, TSP-1 (Thrombospondin 1), Transferrin, or a combination of two or more thereof.

18. The method of any one of claims 15-17, wherein the subject has a history of infertility.

19. The method of any one of claims 15-18, wherein the subject is from 21 to 50 years ofage.

20. The method of any one of claims 15-19, wherein the subject is female.

21. The method of any one of claims 15-20, wherein the subject has a history of one or morefailed in vitro fertilization (IVF) cycles.

22. The method of claim 18, wherein the subject has a lower level of anti-Mullerian hormoneas compared to a subject without a history of infertility.

23. The method of any one of claims 15-22, wherein the subject has premature ovarianfailure (POF).

24. The method of any one of claims 15-23, wherein the subject has polycystic ovarysyndrome (PCOS).

25. The method of any one of claims 15-24, wherein the subject has one or more of thefollowing symptoms selected from the group consisting of: endometriosis, a thyroid disorder, a blocked fallopian tube, a damaged fallopian tube, pelvic inflammatory disease (PID), a high follicle-stimulating hormone (FSH) level, a low estrogen level, a fibroid, a polyp, an autoimmune disorder, a congenital uterine abnormality, a sexually transmitted infection (STI), a hormonal imbalance, a prolactin imbalance, a cortisol imbalance, a gonadotropin-releasing hormone (GnRH) deficiency, inability to conceive, irregular menstrual periods, absent menstrual periods, painful menstrual periods, cramping, heavy or prolonged menstrual bleeding, endometriosis, hormonal fluctuations, mood disturbances, sudden fluctuations in weight, sleep disturbances, and fatigue, or any combination thereof.

26. The method of any one of claims 15-25, wherein the subject is not menopausal or post-menopausal.

27. The method of any one of claims 1-26, wherein the administering comprises intravenousadministration.

28. The method of claim 27, wherein the intravenous administration is carried out over thecourse of 30, 35, 40, 45, 50, 55, or 60 minutes.Attorney Docket No.66309-727.60129. The method of any one of claims 1-28, wherein the composition is prepared by a processcomprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition.

30. The method of any one of claims 1-29, the method comprising preparing the compositionprior to the administering, wherein the preparing the composition occurs by a process comprising: (a) culturing bone marrow mesenchymal stem cells (BM-MSCs) under the following conditions to produce an MSC conditioned media: (i) oxygen tension below 5%; and (ii) culture media having a pH below 7; (b) harvesting the MSC conditioned media; and (c) formulating the MSC conditioned media to produce the composition.

31. The method of claim 29 or claim 30, wherein the culture media is serum-free.

32. The method of any one of claims 29-31, wherein the culture media has a glucoseconcentration below 4.5 g / L.

33. The method of any one of claims 29-32, wherein formulating the MSC conditionedmedia comprises exchanging the conditioned media for a pharmaceutically acceptable formulation.

34. The method of claim 33, wherein the pharmaceutically acceptable formulation comprisessaline.

35. The method of any one of claims 1-34, wherein the composition comprises at least6x1010to 8x1010extracellular vesicles per mL and is administered at a dose of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 ml.

36. The method of claim 35, wherein the dose is administered in combination with normalsaline at a final volume of 100 mL.

37. The method of any one of claims 1-36, wherein the composition comprises saline (0.9%sodium chloride).

38. The method of claim 37, wherein the saline is present in the composition at about 80% toabout 95% saline.

39. The method of any one of claims 1-38, wherein the composition comprises sodiumchloride, sodium lactate, potassium chloride, and calcium chloride.

40. The method of any one of claims 1-39, wherein the molecular weight of any non-excipient component of the composition is greater than about 10 kDa (kilodaltons).

41. The method of any one of claims 1-40, wherein the composition comprises apolysaccharide.Attorney Docket No.66309-727.60142. The method of claim 41, wherein the polysaccharide is present in the composition atabout 0.3 M or about 60 mM.

43. The method of any one of claims 1-42, wherein any non-excipient component of thecomposition has a size of less than about 0.2 microns.

44. The method of any one of claims 1-43, wherein the composition is sterile by USP <71>.

45. The method of any one of claims 1-44, wherein the composition is endotoxin USP <85>free.

46. The method of any one of claims 1-45, wherein the composition is negative formycoplasma DNA.

47. The method of any one of claims 1-46, wherein the composition is cell-free.

48. The method of any one of claims 1-47, wherein the composition is stored between -80 °Cand -60 °C.

49. The method of claim 48, wherein the composition is administered within 6 hours of thawwhen maintained at ambient temperature.

50. The method of any one of claims 1-49, wherein the composition is present in a glass vial.

51. The method of any one of claims 1-50, wherein the composition is formulated forintravenous administration.

52. The method of any one of claims 1-51, wherein the composition has a pH of about 6 toabout 7.5.

53. The method of any one of claims 29-52, wherein the BM-MSCs are negative for CD14,CD31, CD34, and CD45.

54. The method of any one of claims 29-53, wherein the BM-MSCs are positive for CD73,CD105, CD166, and CD90.

55. The method of any one of claims 29-54, wherein the BM-MSCs are capable ofundergoing trilineage differentiation in vitro toward adipocyte, osteoblast, and chondrocyte phenotypes.

56. The method of any one of claims 29-55, wherein the BM-MSCs are obtained from aniliac crest aspiration of a single donor.

57. The method of any one of claims 1-56, wherein a concentration of the one or more EVsin the composition is measured by nanoparticle tracking analysis (NTA).Attorney Docket No.66309-727.60158. The method of claim 57, wherein the NTA comprises light scatter and fluorescenceevaluation.

59. The method of claim 57 or 58, wherein the concentration of the one or more EVs isabout 10 billion to about 250 billion EVs per mL of the composition.

60. The method of any one of claims 57-59, wherein the concentration of the one or moreEVs is at about 1 billion to about 40 billion EVs per ml of the composition.

61. The method of any one of claims 1-60, wherein the one or more EVs have an averagediameter of about 30 nm to about 170 nm.

62. The method of any one of claims 1-61, wherein a total protein concentration of thecomposition is about 10 to about 40 µg per ml of the composition.

63. The method of any one of claims 1-62, wherein a total protein concentration of thecomposition is about 1.5 to about 6 µg per ml of the composition.

64. The method of claim 62 or 63, wherein the total protein concentration is measured byELISA.

65. The method of any one of claims 1-64, wherein the one or more EVs comprise hsa-miR-21-5p, miR-24-3p, hsa-miR-222-3p, hsa-miR-27b-3p, and Let-7.

66. The method of any one of claims 1-65, wherein the one or more EVs comprise TIMP-1(tissue inhibitor of metalloproteinases 1), TIMP-2 (tissue inhibitor of metalloproteinases 2), CD63 antigen, Ferritin, IGFBP-4 (Insulin-like growth factor binding protein-4), and MIF (Macrophage migration inhibitory factor).

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