Regenerative cells and / or derivatives thereof for treatment of female genitourinary conditions, degeneration, and / or damage

Abstract

Discloses herein are methods of treating diseases, disorders, or conditions related to the female genitourinary system. Also disclosed are compositions and delivery devices with therapeutic use in the treatment. Specifically, aspects of the disclosure relate to a degradable and biocompatible device that effectively delivers a therapeutic agent to a subject in need thereof comprising at least one of a stem cell or / and an extracellular vesicle derived thereof, wherein the cells are grown in conditioned medium comprising estrogen or were primed with estrogen, or any combination thereof.

Description

VABIO.002WO2 PCT APPLICATIONREGENERATIVE CELLS AND / OR DERIVATIVES THEREOF FOR TREATMENT OF FEMALE GENITOURINARY CONDITIONS, DEGENERATION, AND / OR DAMAGEINCORPORATION BY REFERENCE TO ANY PRIORITY APPLICATIONS

[0001] Any and all applications for which a foreign or domestic priority claim is identified in the Application Data Sheet as filed with the present application are hereby incorporated by reference under 37 CFR 1.57.

[0002] This application claims priority to U. S. Provisional Application No.63 / 736,204, filed December 19, 2024, U. S. Provisional Application No. 63 / 736,375, filed December 19, 2024, and U. S. Provisional Application No. 63 / 736,303, filed December 19, 2024. Each of the aforementioned applications is incorporated by reference herein in its entirety, and each is hereby expressly made a part of this specification.BACKGROUNDField of the Disclosure

[0003] This application relates generally to the treatment of the female genitourinary tract.Description of Related Art

[0004] Medical conditions, degeneration, abrasions, and other types of damage to the female genitourinary tract are relatively common, impairing quality of life. Causes of such conditions and damage include genitourinary syndrome of menopause (GSM), postpartum vaginal damage (PVD), Mayer -Rokitansky -Kiister -Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), and pelvic floor dysfunction, female infertility caused by refractory lower reproductive tract inflammation, immune imbalance of the uterus, episiotomy, sexual activity, skin conditions, conditions such as endometriosis and interstitial cystitis and polycystic ovary syndrome, human immunodeficiency virus (HIV), vaginal infections, sexually transmitted infections (STIs), hormonal changes, forms of hair removal such as waxing and shaving, complications with foreign objects such as menstrual products, medications, scarring or tissue damage, physical trauma, medical procedures such as radiation therapy, birth control methods,VABIO.002WO2 PCT APPLICATIONcancers and precancerous conditions, and structural abnormalities including uterine fibroids or ovarian cysts.SUMMARY OF CERTAIN INVENTIVE ASPECTS

[0005] Aspects of the present disclosure relate to non-hormonal regenerative therapies. These therapies may in some embodiments include stem cells and / or their derivatives, which offer a safe, long-lasting alternative for women suffering from genitourinary conditions such as Genitourinary Syndrome of Menopause (GSM) — particularly those who cannot use hormonal treatments due to a personal or family history of breast cancer. The treatment of the present disclosure addresses a critical need for the aging female population in seeking effective, hormone-free options due to the potential risks associated with hormone-based therapies. In addition to their clinical benefits, these therapies are also user-friendly. Patients can self-admimster them at home without frequent doctor visits for routine blood hormone monitoring. This convenience enhances patient compliance and quality of life while reducing the economic and social burden on the healthcare system.

[0006] Also disclosed herein is a method of treating a subject having a disease, disorder, or condition, the method comprising administering to the subject a composition comprising an at least one primed stem cell, at least one extracellular vesicle derived from a primed stem cell, or a combination thereof. In some embodiments, the priming of the at least one stem cell comprises at least one of: inducing angiogenesis under hypoxia, administering a hormone, administering a pro- inflammatory molecule, culturing the at least one stem cell in 3D culture, or any combination thereof. In some embodiments, the at least one extracellular vesicle is derived from the stem cell or isolated from a biological fluid. In some embodiments, the biological fluid is a supernatant or the media during the priming step of the stem cell. In some embodiments, the disease, disorder, or condition comprises at least one of: a gender-specific disease, disorder, or condition, genitourinary syndrome of menopause, wound healing, scar treatment, loss of cell elasticity, tissue damage, tissue dryness, tissue irritation, poor circulation, inflammation, infection, or any combination thereof. In some embodiments, the disease, disorder, or condition is female-specific. In some embodiments, angiogenesis is induced by the stem cell undergoing hypoxia. In some embodiments, the pro- inflammatory molecule is a cytokine, preferably wherein the cytokine is IL-6. In some embodiments, the hormone is estrogen.VABIO.002WO2 PCT APPLICATION

[0007] Also disclosed herein is a method of treating a subject having a disease, disorder, or condition associated with the female genitourinary tract, the method comprising administering to the subject a biocompatibie suppository formulated for dissolving inside the vaginal cavity of the subject, the suppository’ comprising a therapeutic composition, the therapeutic composition comprising an at least one primed stem ceil, an at least one extracellular vesicle, or a combination thereof. In some embodiments, the priming of the at least one stem cell comprises at least one of: inducing angiogenesis by hypoxia, administering a hormone, administering a pro- inflammatory molecule, culturing the at least one stem cell in 3D culture, or any combination thereof. In some embodiments, the at least one extracellular vesicle is derived from the stem cell or isolated from a biological fluid. In some embodiments, the biological fluid was the supernatant or media during the priming step of the stem cell. In some embodiments, the at least one primed stem cell is of autologous, allogenic, or xenogeneic origin,

[0008] Disclosed herein is a method of treating a subject having a disease, disorder, or condition associated with the female genitourinary tract. In some embodiments, the method includes administering to the subject a biocompatible suppository formulated for dissolving inside the vaginal cavity of the subject, the suppository comprising a therapeutic composition, the therapeutic composition comprising at least one of: an at least one stem cell, and an at least one extracellular vesicle, wherein the at least one extracellular vesicle is derived from the stem cell or isolated from the stem cell fluid, wherein the at least one stem cell is of autologous, allogenic, or xenogeneic origin, and wherein stem cell is primed to increase efficacy prior to being added to the suppositoiy.

[0009] In some embodiments, the subject is mammalian and / or human. In some embodiments, the disease, disorder, or condition is selected from the group consisting of genitourinary inflammation, damage, dryness, irritation, degeneration, genitourinary syndrome of menopause, postpartum vaginal damage (PVD) Mayer-Rokitansky-Kuster-Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), pelvic floor dysfunction, vaginitis, UTIs, female infertility caused by an immune imbalance in the genital urinary tract, or any combination thereof. In some embodiments, the composition is primed through exposure to an at least one hormone, pro-inflammatory molecule, hypoxia, or any combination thereof.

[0010] In some embodiments, the composition further comprises an antioxidant. In some embodiments, the antioxidant is selected from the group consisting of: ascorbic acid, alphaVABIO.002 WO2 PCT APPLICATIONtocopherol, rutin, quercetin, hesperedin, lycopene, resveratrol, tetrahydrocurcumin, rosmarinic acid, Ellagic acid, chlorogenic acid, oleuropein, alpha-lipoic acid, glutathione, polyphenols, pycnogenol, and a derivative thereof. In some embodiments, the at least one stem cell is a committed progenitor cell, differentiated cell, or any combination thereof. In some embodiments, the stem cell is an embryonic stem cell, cord blood stem cell, placental stem cell, bone marrow stem cell, amniotic fluid stem cell, neuronal stem cell, circulating peripheral blood stem cell, mesenchymal stem cell, mesenchymal stromal cell, germinal stem cell, adipose tissue derived stem cell, exfoliated teeth derived stem cell, hair follicle stem cell, dermal stem cell, parthenogemcally derived stem cell, reprogrammed stem cell, side population stem cell, or any combination thereof,

[0011] In some embodiments, the stem cell is the mesenchymal stromal cell or an induced pluripotent stem cell (iPSC). In some embodiments, the at least one extracellular vesicle is derived from a mesenchymal stem cell and / or a mesenchymal stromal cell. In some embodiments, the at least one extracellular vesicle is derived from a human umbilical mesenchymal stem cell and / or a human umbilical mesenchymal stromal cell. In some embodiments, the at least one extracellular vesicle is isolated from animal blood. In some embodiments, the blood is from a young animal. In some embodiments, the blood is of a young sheep. In some embodiments, the at least one extracellular vesicle is between about 30 nm and about 150 nm in diameter.

[0012] In some embodiments, the at least one stem cell, the at least one extracellular vesicle, or both, comprise a tetraspanin molecule. In some embodiments, the tetraspanin molecule is a CD9, CD63, CD151, Tspan9, or Tspan32. In some embodiments, the at least one stem cell, the at least one extracellular vesicle, or both, comprise at least one of: a heat shock protein, a stress- associated protein, a growth factor receptor, or any combination thereof. In some embodiments, the heat shock protein is selected calreticulin, hsp20, or hsp90. In some embodiments, the stress- associated protein is phosphatidyl serine. In some embodiments, the growth factor receptor is IL-3 receptor, VEGF receptor, CXCL12, or c-met. In some embodiments, the biocompatible suppository comprises chitosan HC1, cross-linking agent, collagen type I, and carboxymethyl cellulose. In some embodiments, the weight ratio of chitosan HC1, cross-linking agent’s collagen type I, and carboxymethyl cellulose is about 40:30:30. In some embodiments, the bioconipatible suppository’ also comprises a mucolytic enzyme. In some embodiments, the mucolytic enzyme is DNase 1.VABIO.002WO2 PCT APPLICATION

[0013] Also disclosed herein is a biocompatible suppository formulated for dissolving inside the vaginal cavity of a subject, the suppository comprising a therapeutic composition with use against a disease, disorder, or condition associated with the female genitourinary tract.

[0014] In some embodiments, the subject is mammalian and / or human. In some embodiments, the therapeutic composition comprises at least one of: an at least one stem cell; and / or an at least one extracellular vesicle, wherein the at least one extracellular vesicle is derived from the stem cell or isolated from the stem cell fluid, wherein the at least one stem cell is of autologous, allogenic, or xenogeneic origin, wherein the composition is primed to increase efficacy prior to being added to the suppository. In some embodiments, the disease, disorder, or condition is selected from the group consisting of genitourinary inflammation, damage, dryness, irritation, degeneration, genitourinary syndrome of menopause, postpartum vaginal damage (PVD) Mayer- Rokitansky-Kuster-Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), pelvic floor dysfunction, vaginitis, UTIs, female infertility caused by an immune imbalance in the genital urinary tract, or any combination thereof.

[0015] In some embodiments, the composition or stem cell is primed through exposure to an at least one hormone, pro- inflammatory molecule, hypoxia, or any combination thereof. In some embodiments, the composition further comprises an antioxidant. In some embodiments, the antioxidant is selected from the group consisting of: ascorbic acid, alpha tocopherol, rutin, quercetin, hesperedm, lycopene, resveratrol, tetrahydrocurcumin, rosmarinic acid, Ellagic acid, chlorogenic acid, oleuropein, alpha-lipoic acid, glutathione, polyphenols, pycnogenol, and a derivative thereof. In some embodiments, the at least one stem cell is a committed progenitor cell, differentiated cell, or any combination thereof.

[0016] In some embodiments, the stem cell is an embryonic stem cell, cord blood stem cell, placental stem cell, bone marrow stem cell, amniotic fluid stem cell, neuronal stem cell, circulating peripheral blood stem cell, mesenchymal stem cell, mesenchymal stromal cell, germinal stem cell, adipose tissue derived stem cell, exfoliated teeth derived stem cell, hair follicle stem cell, dermal stem cell, parthenogemcally derived stem cell, reprogrammed stem cell, side population stem cell, or any combination thereof. In some embodiments, the stem cell is the mesenchymal stem cell, mesenchymal stromal cell, or an induced pluripotent stem cell (iPSC). In some embodiments, the at least one extracellular vesicle is derived from a mesenchymal stem cell and / or a mesenchymal stromal cell. In some embodiments, the at least one extracellular vesicle isVABIO.002WO2 PCT APPLICATIONderived from a human umbilical mesenchymal stem cell and / or a human umbilical mesenchymal stromal cell. In some embodiments, the at least one extracellular vesicle is isolated from animal blood. In some embodiments, the blood is from a young animal. In some embodiments, the blood is of a young sheep.

[0017] In some embodiments, the at least one extracellular vesicle is between about 30 nm and about 150 nm in diameter. In some embodiments, the at least one stem cell, the at least one extracellular vesicle, or both, comprise a tetraspanin molecule. In some embodiments, the tetraspanin molecule is a CD9, CD63, CD151, Tspan9, or Tspan32. In some embodiments, the at least one stem cell, the at least one extracellular vesicle, or both, comprise at least one of: a heat shock protein, a stress-associated protein, a growth factor receptor, or any combination thereof. In some embodiments, the heat shock protein is selected from calreticulin, hsp20, and hsp90. In some embodiments, the stress-associated protein is phosphatidyl serine. In some embodiments, the growth factor receptor is IL-3 receptor, VEGF receptor, CXCL12, or c-met. In some embodiments, the biocompatible suppository comprises chitosan HO, cross-linking agent, collagen type I, and carboxymethyl cellulose. In some embodiments, the weight ratio of chitosan HO, cross-linking agent’s collagen type I, and carboxymethyl cellulose is about 40:30:30. In some embodiments, the biocompatible suppository also comprises a mucolytic enzyme. In some embodiments, the mucolytic enzyme is DNase 1.

[0018] Also disclosed herein is a composition with use against a disease, disorder, or condition associated with the female genitourinary tract. The composition may include, for example, at least one of: an at least one stem cell; and / or an at least one extracellular vesicle, wherein the at least one extracellular vesicle is derived from the stem cell or isolated from the stem cell fluid, wherein the at least one stem cell is of autologous, allogenic, or xenogeneic origin, and wherein the composition or stem cell is primed to increase efficacy prior to being added to the suppositoiy.

[0019] In some embodiments, the disease, disorder, or condition is selected from the group consisting of genitourinary’ inflammation, damage, dryness, irritation, degeneration, genitourinary syndrome of menopause, postpartum vaginal damage (PVD) Mayer-Rokitansky-Kuster-Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), pelvic floor dysfunction, vaginitis, UTIs, female infertility caused by an immune imbalance in the genital urinary' tract, or any combination thereof. In some embodiments, the composition or stem cell isVABIO.002WO2 PCT APPLICATIONprimed through exposure to an at least one hormone, inflammatory molecule, hypoxia, or any combination thereof. In some embodiments, the composition further comprises an antioxidant.

[0020] In some embodiments, the antioxidant is selected from the group consisting of: ascorbic acid, alpha tocopherol, rutin, quercetin, hesperedin, lycopene, resveratrol, tetrahydrocur cumin, rosmarinic acid, Ellagic acid, chi orogenic acid, oleuropein, alpha-lipoic acid, glutathione, polyphenols, pycnogenol, and a derivative thereof. In some embodiments, the at least one stem cell is a committed progenitor cell, differentiated cell, or any combination thereof. In some embodiments, the stem cell is an embryonic stem cell, cord blood stem cell, placental stem cell, bone marrow stem cell, amniotic fluid stem cell, neuronal stem cell, circulating peripheral blood stem cell, mesenchymal stem cell, mesenchymal stromal cell, germinal stem cell, adipose tissue derived stem cell, exfoliated teeth derived stem cell, hair follicle stem cell, dermal stem cell, parthenogenically derived stem cell, reprogrammed stem cell, side population stem cell, or any combination thereof.

[0021] In some embodiments, the stem cell is the mesenchymal stem cell, mesenchymal stromal cell, or an induced pluripotent stem cell (iPSC). In some embodiments, the at least one extracellular vesicle is derived from a mesenchymal stem cell and / or a mesenchymal stromal cell. In some embodiments, the at least one extracellular vesicle is derived from a human umbilical mesenchymal stem cell and / or a human umbilical mesenchymal stromal cell. In some embodiments, the at least one extracellular vesicle is isolated from animal blood. In some embodiments, the blood is from a young animal. In some embodiments, the blood is of a young sheep. In some embodiments, the at least one extracellular vesicle is between about 30 nm and about 150 nm in diameter. In some embodiments, the at least one stem cell, the at least one extracellular vesicle, or both, comprise a tetraspanin molecule. In some embodiments, the tetraspanin molecule is a CD9, CD63, GDI 51, Tspan9, or Tspan32. In some embodiments, the at least one stem cell, the at least one extracellular vesicle, or both, comprise at least one of: a heat shock protein, a stress-associated protein, a growth factor receptor, or any combination thereof.

[0022] In some embodiments, the heat shock protein is selected calreticulin, hsp20, or hsp90. In some embodiments, the stress-associated protein is phosphatidyl serine. In some embodiments, the growth factor receptor is IL-3 receptor, VEGF receptor, CXCL12, or c-met. In some embodiments, the biocompatible suppository comprises chitosan HC1, cross-linking agent, collagen type I, and carboxymethyl cellulose. In some embodiments, the weight ratio of chitosanVABIO.002WO2 PCT APPLICATIONHC1, cross-linking agent’s collagen type I, and carboxymethyl cellulose is about 40:30:30. In some embodiments, the biocompatible suppository also comprises a mucolytic enzyme. In some embodiments, the mucolytic enzyme is DNase 1.BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 illustrates a non-limiting example cartoon schematic of the composition of the present disclosure, comprising at least one of the following: human umbilical cord mesenchymal stromal cells.

[0024] Figures 2A-2C illustrate non-limiting experimental data showing that HUMSCs stimulate the growth of VK2 human vaginal epithelial cells and inhibit apoptosis in vitro. Figure 2A is a non-limiting example cartoon schematic of the experiment, including VK2 cells cocultured with hucMSC, human fibroblasts (HFF-1 ), or KSFM medium in a 0.4-pm pore transwell system, as reported by Zhongyi Zhu et al. at page 250, which has been incorporated by reference in its entirety. Figure 2B is a bar graph displaying cell viability as tested by a formazan reduction assay (CCK-8) after 24 hours of co-culture. Figure 2C is a bar graph illustrating the ratio of apoptosis in the experimental VK2 cell groups, as determined by flow cytometry. Data are presented as mean ± SD, N::::3; *, P < 0.05; **, P < 0.01; ***ty p < 0.0001.

[0025] Figures 3A-3B illustrate non-limiting experimental data showing that HUMSC administration increases collagen expression in the anterior vaginal wall of menopausal rats. In a paper by Kadek Fajar Marta, et al. at page 88, which has been incorporated by reference in its entirety, the histochemical score (H-score) of the samples m the HUMSC treatment group was shown to be higher than the H-score in the control group. Figure 3A depicts two non-limiting photographs of the anterior vaginal wall of experimentally treated menopausal rats. Higher staining is correlated with higher collagen expression. The darker arrows represent areas of high staining while the lighter arrows represent areas of lower staining. The treatment group had higher staining overall than the control group, with an H-score of 4.3, compared to the control group’s H-score of 1.37. Figure 3B shows a chart comparing the collagen expression levels in the treatment and control groups of rats. In Figure 3B, the number of rats in both the treatment group and control group is indicated out of 18 rats. In the treatment group, 13 out of 18 rats showed high collagen expression, compared to 0 out of 18 rats in the control group.VABIO.002WO2 PCT APPLICATION

[0026] Figures 4A-4B illustrate non-limiting experimental data showing that secreted exosomes derived from HUMSCs stimulate the growth of a VK2 cell line in a dose-dependent manner, as discussed in a paper by Zhongyi Zhu et al. (page 254), which has been incorporated by reference in its entirety. Figure 4A is a bar graph illustrating the measurement of VK2 cells at various concentrations of exosomes derived from hucMSC (hucMSC-Ex). In Figure 4A, cell viability, as measured by a tetrazolium reduction assay using a Cell-Counting Kit-8 (CKK-8), was shown to increase as the concentration of hucMSC-Ex treatment increased in VK2 cell samples. Figure 4B is a bar graph illustrating the cell apoptosis ratio among VK2 cells treated with hucMSC-Ex compared to cells treated with HFF-1 exosomes and cells treated with keratmocyte serum-tree medium (KSFM). The cell apoptosis ratio among VK2 cells treated with hucMSC-Ex. was shown to be lower than in VK2 cells treated with either human fibroblast (HFF-1) exosomes or keratinocyte serum-free medium (KSFM).

[0027] Figures 5A-5C illustrate non-limiting experimental data demonstrating that Human Umbilical Cord Mesenchymal Stem Cells-derived Exosomes (ucMSC-Ex) increase VK2 cell viability, proliferation, and migration in vitro, as discussed in a paper by Tao Zhang, et al. on page 14295, which has been incorporated by reference in its entirety. Figure 5A is a bar graph illustrating the increasing viability of VK2 cells at varying concentrations of ucMSC-ex, as determined by a Cell Counting Kit-8 (CCK-8) assay. Figure 5B is a bar graph showing a quantification of S-phase cell proportion. In Figure 5B, cells treated with ucMSC-ex showed higher proliferation as compared to cells treated with PBS. Figure 5C is a bar graph quantifying the cell migration rate of cells treated with either ucMSC-ex or phosphate-buffered saline (PBS) after 18 hours and 36 hours. ucMSC-ex-treated cells showed a higher cell migration rate as compared to PBS-treated cells when measured after both 18 hours and 36 hours.

[0028] Figures 6A-6D illustrate non-limiting experimental data of vaginal epithelial morphology following treatment, as discussed in a paper by Tao Zhang, et al. on pages 14297-98, which has been incorporated by reference in its entirety. Figure 6A depicts non-limiting example images of vaginal epithelial morphology after hematoxylin and eosin (HE) staining among rats with no treatment or operations (Sham), rats with ovariectomy, ovariectomized rats with a hydrogel treatment, ovariectomized rats with exosome-hydrogel treatment, and ovariectomized rats with estrogen treatment. Figure 6B presents non-limiting example images of vaginal epithelial morphology after HE staining for the same experimental rat groups, illustrating the average opticalVABIO.002WO2 PCT APPLICATIONdensity analysis of Ki-67 and CD 31 expression in the vaginal epithelium and lamina propria. Figure 6C illustrates a non-limiting example of a bar graph quantifying Ki-67 staining, as measured by optical density (OD). Data are presented as mean ± SD, N = 6; *, P < 0.05; **, P < 0.01; ***, P < 0.001. Figure 6D illustrates a non-limiting example bar graph for CD 31 staining, as determined by optical density (OD). Data are presented as mean ± SD, N = 6; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

[0029] Figures 7A-7G illustrate non-limiting example experimental data showing that human umbilical cord mesenchymal stromal cell-derived exosomes (HucMSC-Exo) promote the viability, contractility, and production of collagen in human primary vaginal fibroblasts in vitro, as further discussed in a paper by Lei-Mei Xu, et al, on pages 719, 721, and 722, which has been incorporated by reference in its entirety. Figure 7 A illustrates a non-limiting example line graph displaying the effect of human umbilical cord mesenchymal stromal cell-derived exosome (hucMSC-Exo) on the viability of fibroblasts from 0 hours to 72 hours in varying concentrations of hucMSC-Exo, as measured by a Cell Counting Kit-8 (CCK-8) assay. Figure 7B illustrates a non-limiting example bar graph comparing cell viability data among the four treatment groups with different hucMSC-Exo concentrations at the 48-hour time point. Figure 7C is a non-limiting representative image of gel contraction following incubation with hucMSC-Exo (4 or 6 gg / mL) for 48 hours. Figure 7D illustrates a non-limiting example bar graph illustrating the quantification of gel contraction after incubation with hucMSC-Exo (4 or 6 gg / mL) for 48 hours. Gel contraction was measured as a percentage of the area of the control group. Figure 7E displays non-limiting example images of protein expression of Coll, as determined by immunofluorescence. A Sirius Red collagen staining kit was used to determine the total secreted collagen level in the fibroblasts’ supernatant. Figure 7F illustrates a nonlimiting example bar graph displaying the total secreted collagen (in gg / mL) for the control group, the group treated with 4 gg / mL of exosomes, the group treated with 6 gg / mL of exosomes, and the group treated with 10% fetal bovine serum as an additional control. Figure 7G illustrates a non-limiting example bar graph displaying the mean fluorescence activity for the control group, the group treated with 4 gg / mL of exosomes, the group treated with 6 gg / mL of exosomes, and the group treated with 10% fetal bovine serum as an additional control. The data were analyzed by one-way ANOVA (means ± SD).aP < 0.05;bP < 0.01; °P< 0.001;dP< 0.0001.VABIO.002WO2 PCT APPLICATION

[0030] Figures 8A-8C illustrate non-limiting experimental data and graphics characterizing microRNA profiles in human umbilical cord mesenchymal stromal cell-derived exosomes (HucMSC-Exo), as further discussed in a paper by Lei-Mei Xu et al. on page 724, which is incorporated by reference in its entirety. Additionally, this experimental data confirms the downregulation of MMPl 1 as a beneficial target for microRNA in enhancing collagen expression in human primary vaginal fibroblasts. See id. Figure 8A is a non-limiting example pie chart showing the variety' and proportion of different miRNAs in hucMSC-Exo. Figure 8B is a non¬ limiting example schematic for a regulatory network illustrating the top predicted microRNAs (miRNAs) associated with the gene of interest, matrix metalloproteinase 11 (MMP11), and the enrichment terms of these miRNAs in the Gene Ontology analysis. Figure 8C is a non-limiting bar graph showing the mRNA expression of MMP11 as a proportion of the control group’s mRNA expression. Quantitative real-time polymerase chain reaction was performed to evaluate the mRNA levels of MMPl 1,bP < 0,01.DETAILED DESCRIPTION OF CERTAIN INVENTIVE EMBODIMENTS

[0031] Aspects of the present disclosure relate to the treatment of a disease, disorder, or condition linked to the female genitourinary system. In some embodiments, the conditions comprise degeneration and / or damage to the genitourinary system. In some embodiments, the treatment comprises administering to a subject in need thereof at least one regenerative cell, at least one derivative of a regenerative cell, or a combination thereof. In some embodiments, the at least one regenerative cell comprises a natural stem cell, such as a mesenchymal stem cell, mesenchymal stromal cell (MSC), or an induced pluripotent stem cell (IPSC). In some embodiments, the IPSC, a type of pluripotent stem cell generated from adult cells using reprogramming technology, can be isolated from patient-specific cell lines. This avoids any immunogenicity issues and is also a good resource for IPSC-derived culture medium (IPSC-CM) and extracellular vesicles (IPSC-EV). In some embodiments, the at least one derivative of the regenerative cell comprises an extracellular vesicle, such as an exosome, microvesicle, or an apoptotic body.

[0032] In some embodiments, at least 1, 2, or 3 of: a mesenchymal stromal cell, a mesenchymal stromal cell culture medium (MSCs-CM), and a mesenchymal stromal cell-derived extracellular vesicle (MSCs-EV), or any combination thereof, are administered to the subject toVABIO.002WO2 PCT APPLICATIONtreat a genitourinary condition. In some embodiments, the mesenchymal stromal cell is a human umbilical mesenchymal stromal cell (HUMSC). In some embodiments, the mesenchymal stromal cell culture medium is human umbilical mesenchymal stromal cell culture medium (HUMSCs-CM). In some embodiments, the mesenchymal stromal cell-derived extracellular vesicle is a human umbilical mesenchymal stromal cell-derived extracellular vesicle (HUMSCs-EV).

[0033] In some embodiments, induced pluripotent stem cell-derived culture medium (iPSC-CM) and extracellular vesicles (EVs) are used individually or in combination for genitourinary treatment.

[0034] In some embodiments, a mesenchymal stromal cell, an iPSC-derived extracellular vesicle (EV), or a combination thereof, is used to treat a genitourinary condition. In some embodiments, the EV is derived from a mesenchymal stromal cell as an extracellular vesicle (an MSC-EV). In some embodiments, the extracellular vesicle is derived from a human umbilical mesenchymal stromal cell (HUMSC) and is referred to as a HUMSC-EV.

[0035] In some embodiments, animal blood is used to isolate the extracellular vesicles (EVs), which are used in a genitourinary treatment. In some embodiments, young animal blood is used to isolate EV, which has use in the treatment. In some embodiments, young sheep blood is used to isolate EVs, which have use in the treatment. In some embodiments, the purified plasma of young animals is used to isolate EVs, which have use in the treatment.

[0036] In some embodiments, animal blood is used to isolate small extracellular vesicles (sEVs), which are used in a genitourinaiy treatment. In some embodiments, blood from young animals is used to isolate sEVs, which are then utilized for therapeutic purposes. In some embodiments, young sheep blood is used to isolate sEVs, which are used in a treatment. In some embodiments, the purified plasma of young animals is used to isolate sEVs, which are used in a treatment

[0037] In some embodiments, the disease, disorder, or condition linked to the female genitourinary system comprises at least one of: vaginal dryness or damage, genitourinaiy syndrome of menopause (GSM), postpartum vaginal damage (PVD), Mayer-Rokitansky-Kuster-Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), pelvic floor dysfunction, female infertility caused by refractory lower reproductive tract inflammation, immune imbalance of the uterus, episiotomy, sexual activity, skin conditions, conditions such as endometriosis and interstitial cystitis and polycystic ovary syndrome, human immunodeficiencyVABIO.002WO2 PCT APPLICATIONvirus (HIV), vaginal infections, sexually transmitted infections (STIs), hormonal changes, forms of hair removal such as waxing and shaving, complications with foreign objects such as menstrual products, medications, scarring or tissue damage, physical trauma, medical procedures such as radiation therapy, birth control methods, cancers and precancerous conditions, and structural abnormalities including uterine fibroids or ovarian cysts.

[0038] Also disclosed herein is a method of treating the disease, disorder, or condition linked to the female genitourinary system in a subject in need thereof, the method comprising administering to the subject a therapeutic agent effective against the disease, disorder, or condition. In some embodiments, the therapeutic agent comprises an at least one regenerative cell, optionally in combination with conditioned media and / or extracellular vesicles. In some embodiments, the regenerative cell is a stem cell or a stem cell derivative. In some embodiments, the regenerative cell is administered to the subject by being loaded into a suppository or a suppository -type device. In some embodiments, the vaginal suppository or suppository-type device comprises a biocompatible gel. In some embodiments, the vaginal suppository has a roughly conical or cylindrical shape.

[0039] In some embodiments, the therapeutic agent is administered to the subject via suppositories, gels, creams, lotions, ointments, foams, wipes, lubricants, irrigations, hydrotherapies, capsules, tablets, pellets, microparticles, nanoparticles, patches, films, or rings. In some embodiments, the therapeutic agent is administered by a biocompatible vaginal suppository. In some embodiments, the biocompatible vaginal suppository comprises a scaffold made primarily from chitosan HQ, a cross-linking agent, collagen type I, and carboxymethyl cellulose. In some embodiments, a biocompatible vaginal suppository comprises theobroma oil or cocoa butter. In some embodiments, a biocompatible vaginal suppositoiy comprises synthetic triglycerides. In some embodiments, a biocompatible vaginal suppository comprises glycerinated gelatin. In some embodiments, a biocompatible vaginal suppositoiy comprises a polyethylene glycol polymer. In some embodiments, a biocompatible vaginal suppository comprises surfactants or preservatives.Genitourinary Syndrome of Menopause (GSM)

[0040] Genitourinary syndrome of menopause (GSM) describes the multiple symptoms resulting from estrogen deficiency in the female genitourinary tract, including the vagina, labia, urethra, and bladder. This syndrome includes genital symptoms of dryness, burning,VABIO.002WO2 PCT APPLICATIONand irritation; urinary symptoms of dysuria, urgency, and recurrent urinary tract infections (UTIs); and sexual symptoms of pain and dryness. Vulvovaginal atrophy is a component of GSM.

[0041] GSM is typically diagnosed by identifying some of the aforementioned symptoms in a patient, either through a discussion of the patient's history or a physical examination. GSM most commonly develops in the setting of hypoestrogenism. Therefore, patients at risk of GSM are typically postmenopausal women, since menopause is associated with a hypoestrogenic state. However, GSM can also occur in individuals with other conditions associated with low estrogen, including those who have undergone surgical menopause following bilateral oophorectomy (with or without hysterectomy), or those who have used cancer treatments such as surgery', pelvic radiation therapy, gonadotropin-releasing hormone agonists or aromatase inhibitors (AIs), or chemotherapy. Breast cancer survivors, particularly, are at increased risk due to treatments such as chemotherapy or aromatase inhibitors, which can contribute to the early onset of GSM symptoms. A dditionally, GSM is a risk for pati ents who have experienced primary ovarian insufficiency (POI), hypothalamic amenorrhea, or who are breastfeeding. Additional risk factors include ovarian failure due to radiation or arterial embolization, hypothalamic-pituitary disorders, and the use of antiestrogen medications such as leuprolide or gonadotropin-releasing hormone antagonists, often prescribed for endometriosis.

[0042] In contrast to vasomotor symptoms (VMS) that usually improve over time, GSM is generally progressive without effective therapy. GSM affects approximately up to 84% of postmenopausal women and can significantly impair health, sexual function, and quality of life. Current available treatments for GSM include: 1) Vaginal lubricants and moisturizers, which must be used frequently (several times per week), have veiy little clinical support for their efficacy, and can often be hyperosmolar in the case of water-based products, potentially leading to epithelial cellular toxicity, damage in cultures of epithelial cells, and ectocervical explants; 2) Vaginal estrogens and dehydroepiandrosterone (DHEA), which often entail the same adverse effects associated with other vaginal estrogen therapies, including vaginal discharge, vulvovaginal candidiasis, vaginal bleeding, and breast pain; 3) Systemic hormone therapy and the estrogen agonist / antagonist ospemifene, which carry risks such as breast and endometrial cancer, and cardiovascular disease (CVD); and 4) Energy-based therapies, including laser and radio-frequency devices, which have not yet been approved for effectiveness and safety.VABIO.002WO2 PCT APPLICATION

[0043] To fulfill this unmet need in women’s health and improve female life quality, there is an urgent need for a non-hormonal, non-invasive, safe, convenient, and efficient therapy to treat GSM.Postpartum Vaginal Damage (PVD)

[0044] Postpartum vaginal damage (PVD) affects up to 90% of women who give birth through vaginal delivery. PVD commonly causes tears to the vagina and the surrounding perineum, caused by stretching during childbirth. There are four degrees of postpartum vaginal damage. First-degree tears are the least severe and involve tears to the perineum and urethra. Second-degree tears are the most common form of PVD and are more severe, as they may go deeper into the vagina and typically require stitches. Third-degree tears extend from the vagina to the anus and damage the anal sphincter muscles. Finally, fourth-degree tears are the least common and the most severe, and extend from the vagina, through the perineal area and anal sphincter muscles, into the rectum,

[0045] Phenotypes associated with PVD include perineal trauma, pelvic floor muscle injuries, connective tissue remodeling, nerve injuries, and vascular dysfunction. Perineal trauma can consist of persistent pain, dyspareunia, and pelvic floor disorders. Pelvic floor muscle injuries occur due to overstretching and damage to the levator am muscle complex, particularly the pubococcygeus muscle, which is the shortest and most medial component. During childbirth, the connective tissue elements of the pelvic floor are at risk of stretching and detaching from their bony attachments. Following delivery, there is substantial remodeling of the connective tissue components, characterized by an increase in the synthesis of collagen and elastin. However, the resulting healed connective tissue is not as strong as the original tissue that it replaces. Further, stretching and compression injury to the pudendal nerve has been reported in 38-42% of vaginal deliveries. At one month postpartum, a small but significant vasoconstrictive phenotype has been demonstrated due to an underlying loss of basal nitric oxide contribution.

[0046] PVD is typically diagnosed by a physical exam, assessment of symptoms, and / or imaging techniques. Common symptoms include pain in the vagina or perineum, or going down one’s legs, changes to bowel or bladder habits, or a heaviness or dragging sensation in one’s vagina. Those with PVD are typically females who have given birth vaginally. Those at risk of PVD are pregnant women and women who have recently given vaginal birth. Certain factors can increase the risk of PVD, including nulliparity (first-time mothers), operative vaginal deliveryVABIO.002WO2 PCT APPLICATION(forceps or vacuum), midline episiotomy, advanced maternal age, Asian ethnicity, increased fetal weight, malpresentation — including persistent occiput posterior position and advancing gestational age — and prolonged labor.

[0047] Currently, available treatments for PVD are limited to stitches and home remedies such as applying an ice pack to the affected area, submerging the pelvis in a warm bath, sitting on a donut-shaped pillow, and ingesting over-the-counter medication. PVD can take weeks to heal, and improper care may result in infection.

[0048] Considering the frequency of PVD, the potential severity of damage, and the lack of treatment options, there is an urgent need for a non-hormonal, non-invasive, safe, convenient, and efficient therapy to treat PGD.Mesenchymal Stromal Cells (MSCs)

[0049] Mesenchymal stem cells — a group of adult pluripotent stem cells with the ability to self-renew and differentiate — can be obtained from almost all human tissues. Mesenchymal stromal cells (MSCs) are mesenchymal stem cells that are isolated from a sample, and may comprise a mixed population of stem-like and non-stem cells. In recent years, MSCs have become a research area of high activity due to their wide range of sources, low immunogenicity, and multiple functions based on: I) MSCs’ ability to convey several cell adhesion molecules and receptors, including CXCL12 and integrins, which can induce specialized migration of MSCs to target tissues for an extended period of time; 2) MSCs’ production of MSC-derived bioactive factors known as secretome, which comprise proteins such as IGF, VEGF, FGF2, EGF and HGF cytokines, nucleic acids including micro-RNAs and long non-coding RNAs, and extracellular vesicles (EVs) including exosomes, microvesicles, and microparticles, which modify the microenvironment, thereby restoring organ function; 3) the ability of MSCs to promote proliferation and regulate autophagy and oxidative stress, reduce apoptotic and fibrotic effects to the local cells, and restore the function of the tissues and organs; 4) MSCs’ ability to exert immunomodulatory activities by migrating to affected tissues to inhibit local inflammation without showing systemic effect, thus neither causing systemic suppression of the immune system nor increasing tumor growth; and 5) MSCs’ ability to differentiate into multiple tissue-specific cell types to repair the damaged tissue.VABIO.002WO2 PCT APPLICATION

[0050] Umbilical cord mesenchymal stromal cell-derived exosomes (ucMSC-ex) boast functional similarities with their parent cells and offer several advantages, including a smaller size, fewer side effects, and no ethical dilemmas. This positions them as a promising avenue for tissue regeneration therapy. Exosomes, nano-membrane vesicles of 30-150 nm, are encapsulated within a double membrane and secreted by living cells. They are laden with proteins, mRNAs, miRNAs, and lipids, enabling them to shuttle between cells and deliver biologically active substances to receptor cells, initiating a biological response. The regenerative abilities of exosomes derived from umbilical cord mesenchymal stromal cells are well -documented, including their roles in enhancing functional repair of the spinal cord, promoting ovarian cell proliferation, and boosting angiogenesis. As a major component of paracellular secretion, they are believed to effectively regenerate defective vaginal mucosa.

[0051] Mesenchymal stromal cell (MSC)-based therapy has emerged as a promising approach in regenerative medicine due to the cells’ key properties, including self-renewal, multipotency, and immunomodulatory effects. Among various sources of MSCs, such as bone marrow, adipose tissue, and Wharton’s jelly, human umbilical cord-derived MSCs (hucMSCs) are particularly attractive, as they are typically considered medical waste and can be collected non-mvasively, without ethical concerns. This makes hucMSCs a safe, cost-effective, and scalable cell source. Furthermore, hucMSCs exhibit strong cytokine secretion capabilities and have been widely used in vivo and in vitro tissue repair studies. Nevertheless, several unresolved challenges ------such as progeny cell heterogeneity, potential tumorigenicity, and the risk of immune rejection — continue to limit their clinical application.

[0052] As key secretory mediators of hucMSCs, exosomes have attracted growing interest as promising cell-free therapeutic agents. These nanosized (typically about 30-200 nm), cup-shaped extracellular vesicles possess a lipid bilayer membrane and carry various bioactive molecules, and bypass many risks associated with cell-based therapies. Exosomes are enriched with proteins, mRN As, microRNAs (miRNAs), and lipids, allowing them to mediate intercellular communication by delivering bioactive molecules to recipient cells and triggering specific biological responses. The regenerative potential of exosomes derived from umbilical cord mesenchymal stromal cells is well-documented, with demonstrated roles in promoting spinal cord repair, stimulating ovarian cell proliferation, and enhancing angiogenesis.VABIO.002WO2 PCT APPLICATION

[0053] Human umbilical cord MSC-derived exosomes (hucMSC-Exo) play a significant role in promoting the proliferation of human vaginal epithelial VK2 cells by accelerating the cell cycle and inhibiting apoptosis via exosomal microRNAs in vitro. hucMSC-Exo is also capable of restoring the growth and function of primary fibroblasts from patients with pelvic organ prolapse (POP) by enhancing cell proliferation and increasing collagen I (Coll) expression in vitro. The exosomes, enriched with microRNAs, also downregulated the expression of matrix metalloproteinase 11 (MMP11) to enhance the collagen production (Figures 8A-8C). Furthermore, hucMSC-Exo stimulates both the proliferation and migration of human vaginal epithelial VK2 cells in vitro. At the same time, short-term topical application of an exosome-containing hydrogel modestly increases vaginal epithelial thickness in vivo in a menopause rat model. Also, administration of huMSC resulted in an increase in collagen levels in the anterior vaginal tissue of postmenopausal female rats. In light of this, hucMSCs and their derived exosomes (hucMSC-Ex) could provide safe, effective, and non-hormonal therapy for various conditions in women's health, including genitourinary syndrome of menopause (GSM), postpartum vaginal damage (PVD), Mayer -Rokitansky -Kiister -Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), and pelvic floor dysfunction, female infertility caused by refractory lower reproductive tract inflammation, immune imbalance of the uterus, episiotomy, sexual activity, skin conditions, conditions such as endometriosis and interstitial cystitis and polycystic ovary syndrome, human immunodeficiency virus (HIV), vaginal infections, sexually transmitted infections (STIs), hormonal changes, forms of hair removal such as waxing and shaving, complications with foreign objects such as menstrual products, medications, scarring or tissue damage, physical trauma, medical procedures such as radiation therapy, birth control methods, cancers and precancerous conditions, and structural abnormalities including uterine fibroids or ovarian cysts.hucMSCs

[0054] Human umbilical cord mesenchymal stromal cells (HucMSCs) are the most abundant MSCs and can be obtained from discarded umbilical cords. HucMSCs can secrete chemokines, growth factors, and hormones to affect adjacent cells, which is known as the paracrine effect. The paracrine secretion of HucMSCs plays a crucial role in angiogenesis, anti-VABIO.002WO2 PCT APPLICATIONinflammation, immunomodulation, anti-apoptosis, and anti-fibrosis, thereby improving the microenvironment and promoting the recovery of injured tissues.

[0055] There are several predicted advantages to using HucMSCs compared to other stem cells, including: 1) a higher secretion of paracrine factors; 2) a greater proliferative capacity; 3) weaker immunosuppressive function; 4) no evidence of direct immune rejection in vivo,' and 5) prolonged survival following transplantation into the tissue. This is supported by several papers, which have independently demonstrated the beneficial properties of hucMSCs (Figures 2A-2C, 3 A-3B, 4A-4B, 5A-5C, 6A-6D, and 7A-7G).

[0056] Human umbilical cord MSC extracellular vesicles (HUcMSCs-EVs) are small vesicles with a diameter of <1 pm carrying membrane and cytoplasmic constituents of cellular origin. These vesicles can transfer proteins, mRNA, and bioactive lipids to target cells via surface-expressed ligands and receptors, thereby affecting the phenotype and function of the target cells. EV transplantation is a novel cell-free therapeutic strategy for various degenerative disorders, aiming to achieve therapeutic effects on anti-aging and tissue repair, and is a preferable approach in the clinic.

[0057] In some embodiments, human umbilical cord MSC culture medium (HUMSCs-CM) is used alone or in combination with human umbilical cord MSC extracellular vesicles (HUcMSCs-EVs) for the treatment of genitourinary diseases and disorders such as GSM (Figure 1). HUMSCs, HUMSCs-CM, and HUMSCs-EVs each exhibit paracrine effects that ameliorate ovarian senescence. Regulation of Bax, Bcl-2, and caspase-3 helps to inhibit apoptosis. VEGF, HGF, IGF-1, angiopoietin, and angiogenin play important roles in angiogenesis. Thl / Th2 cytokines play important roles in a balanced local immune response. The AMPK / mTOR signaling pathway plays an important role in the antioxidant process. The mechanism of antifibrosis is not clear.Non-limiting Example Methodology for Cell Priming

[0058] In some embodiments of the present disclosure, the stem cell is primed. In some embodiments, the priming comprises exposing the stem cell to an at least one hormone, pro- inflammatory molecule, an angiogenesis-inducing condition, or any combination thereof. In some embodiments, the angiogenesis-inducing condition comprises hypoxia. In some embodiments, the hormone comprises estrogen. In some embodiments, the inflammatory molecule comprises aVABIO.002WO2 PCT APPLICATIONcytokine, optionally wherein the cytokine is IL-6. In some embodiments, the extracellular vesicle is isolated from the stem cell after priming. In some embodiments, the isolated extracellular vesicle retains the benefits of the primed stem cell or the fluid in which the stem cell was primed, and can thus be referred to as a “primed extracellular vesicle.”

[0059] Also disclosed herein is a therapeutic use for the stem cell and / or extracellular vesicle in treating a disease, disorder, or condition in a subject. In some embodiments, the stem cell and / or extracellular vesicle are primed differently based on the disease, disorder, or condition. Non-limiting examples of priming conditions based on the diseases, disorders, or conditions to be treated are as shown in Table 1.Table 1: Nonlimiting Example Priming Conditions for Treating DiseasesDisease, Disorder, or Condition: Priming Type:General therapy Hormonal treatment (for example: estrogen, Gender-specific diseases androgen, testosterone, progesterone, relaxin, Menopause GnRH, FSH, LH, or hCG)Tissue damage (such as vaginal or urethral) Induce angiogenesis (for example: through Wound healing or scar treatment hypoxia)Poor circulationInflammation (including autoinflammation) Pro-Inflammatory molecule treatment (for Immune disorder example: IL-6, IFN-y, IL- la, IL-ip, IL-25, IL- Infection (such as vaginal or urethral) 6, TNF-a, IL23 or IL- 17)Vaginitis

[0060] In some embodiments, the subject has a disease, disorder, or condition as presented in Table 1 and the stem cell and / or vesicle is primed under more than one condition. In some embodiments, the subject has more than one disease, disorder, or condition. In this case, a cell or vesicle may be primed under more than one condition. In some embodiments, the cell or vesicle is primed through exposure to estrogen, and at least one more priming condition.

[0061] In some embodiments, the cell is primed through hormonal treatment, wherein the hormonal treatment comprises administering an at least one hormone to the cell. In some embodiments, the hormone is mammalian and / or human. In some embodiments, the cell is a stemVABIO.002WO2 PCT APPLICATIONcell. In some embodiments, the extracellular vesicle is later extracted from the cell-culture medium, or the fluid of the cell. In some embodiments, the at least one hormone is selected from: Estrogen, Progesterone, Follicle-Stimulating Hormone (FSH), Luteinizing Hormone (LH), Human Chorionic Gonadotropin (hCG), GnRH (Gonadotropin-Releasing Hormone), androgen, relaxin, or a derivative thereof. In some embodiments, the at least one hormone is added to a cell at a concentration that is about 1 ng / mL, about 1 mg / mL, or any concentration that is between about 1 ng / mL and about 1 mg / mL. In some embodiments, the at least one hormone is added to a cell at a concentration that is about 1 ug / mL, about 100 ug / mL, or any concentration that is between about 1 ug / mL and about 100 ug / mL. In some embodiments, the at least one hormone is administered to the cell for at least about 30 minutes. In some embodiments, the at least one hormone is administered to the cell for about 1 hour, about 12 hours, about 24 hours, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 2 weeks, about 3 weeks, about a month, or any value that is between an hour and a month. In some embodiments, the at least one hormone is administered to the cell for about 1 day, about 3 days, or any value that is between about 1 day and about 3 days. In some embodiments, the cell is washed following hormonal treatment.

[0062] In some embodiments, the cell is primed to increase its immunomodulation activity, wherein the priming comprises administering at least one pro- inflammatory molecule to the cell. In some embodiments, the pro-inflammatory molecule is mammalian and / or human. In some embodiments, the cell is a stem cell. In some embodiments, the extracellular vesicle is later extracted from the cell-culture medium, or the fluid of the cell. In some embodiments, the at least one pro-inflammatory molecule is selected from: interferon gamma (IFN-y), interleukin (IL)- 1 a / p, IL-25, IL-6, TNF-a, IL-17, IL-23, or a derivative thereof. In some embodiments, the at least one pro- inflammatory molecule is added to a cell at a concentration that is about 0.1 ng / mL, about 1 mg / mL, or any concentration that is between about 0.1 ng / mL and about 1 mg / mL. In some embodiments, the at least one pro-inflammatory molecule is added to a cell at a concentration that is about 10 ng / mL, about 100 ng / mL, or any concentration that is between about 10 ng / mL and about 100 ng / mL. In some embodiments, the at least one pro-inflammatory molecule is administered to the cell for at least about 30 minutes. In some embodiments, the at least one pro-inflammatory molecule is administered to the cell for about 1 hour, about 12 hours, about 24 hours, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 2 weeks,VABIO.002WO2 PCT APPLICATIONabout 3 weeks, about a month, or any value that is between an hour and a month. In some embodiments, the at least one pro-inflammatory molecule is administered to the cell for about 1 day, about 2 days, or any value that is between about 1 day and about 2 days. In some embodiments, the cell is washed following pro-inflammatory molecule treatment.

[0063] In some embodiments, the cell is primed to induce or increase angiogenesis, wherein the priming comprises reducing the oxygen content surrounding the cell. In some embodiments, the cell is a stem cell. In some embodiments, the extracellular vesicle is later extracted from the cell culture medium, or the fluid of the cell. In some embodiments, the cell is placed in an atmosphere that comprises about 0.5%, about 7%, or any value between about 0.5% and about 7%, total oxygen content. In some embodiments, the cell is placed in an atmosphere that comprises about 1%, about 2%, or any value between about 1% and about 2%, total oxygen content. In some embodiments, the cell is placed in an atmosphere that comprises about 1 %, about 2%, about 3%, about 4%, about 5%, bout 6%, about 7%, about 8%, about 9%, about 10%, or any value that is between about 1% and about 10%, total carbon dioxide content. In some embodiments, the cell is placed in an atmosphere that comprises balanced nitrogen. In some embodiments, the cell is placed in an atmosphere that comprises sufficiently low oxygen as to induce hypoxia. In some embodiments, the total oxygen content in the atmosphere is reduced through the use of a modular incubator chamber. In some embodiments, the oxygen content is reduced for at least about 30 minutes. In some embodiments, the oxygen content is reduced for about 1 hour, about 12 hours, about 24 hours, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 7 days, about 2 weeks, about 3 weeks, about a month, or any value that is between an hour and a month. In some embodiments, the oxygen content is reduced for about 0.5 days, about 2 days, or any value that is between about 0.5 days and about 2 days. In some embodiments, the cell is returned to normal oxygen levels following hypoxia.

[0064] In some embodiments, the cell is primed to increase trophic and / or immunomodulatory function. In some embodiments, the cell is primed m 3D culture. In some embodiments, the 3D culture generates cell spheroids, mimicking the in vivo niche environment and enhancing the phenotypic profile of cells compared with 2D cultures. In some embodiments, the cell is a stem cell. In some embodiments, stem cell spheroids are generated. In some embodiments, the extracellular vesicle is later extracted from the cell culture medium, or the fluid of the cell.VABIO.002WO2 PCT APPLICATION

[0065] In some embodiments, the cell is primed at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times. In some embodiments, the cell is primed through the same class (i.e. hormonal, pro-inflammatory', hypoxia, or 3D culture) at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 times. In some embodiments, the cell is primed through 1, 2, 3, or 4 of: hormonal, pro-inflammatory, hypoxia, or 3D culture treatment. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, or 9 hormones are administered to the cell. In some embodiments, at least 1, 2, 3, 4, 5, 6, 7, or 9 pro-inflammatory molecules are administered to the cell.Non-limiting Example Methodology for Extracellular Vesicle Isolation

[0066] In some embodiments, the extracellular vesicle is isolated from the stem cell culture medium and / or biological fluid surrounding the stem cell. In some embodiments, the extracellular vesicle is isolated after the stem cell is primed at least once. In some embodiments, the extracellular vesicle is isolated through centrifugation, filtration, pull-down assay, chromatography, or any combination thereof. In some embodiments, the extracellular vesicle is isolated through sequential centrifugation. In some embodiments, the extracellular vesicle is isolated through Tangential flow filtration (TFF) and / or Size-exclusion chromatography (SEC).

[0067] In some embodiments, the presence of the extracellular vesicle in the composition is assessed by quantifying at least one property associated with the extracellular vesicle. In some embodiments, the property is selected from: particle size, concentration, protein content, marker expression, sterility, potency, endotoxin testing, residual DNA / RNA, or any combination thereof. In some embodiments, the presence of the extracellular vesicle in the composition is assessed by at least one of: NTA, TRPS, Western blot, ELISA, nanodrop, activity assay, or any combination thereof. In some embodiments, the composition is adjusted to comprise a therapeutic concentration of the extracellular vesicles.Terms

[0068] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents, applications, published applications, and other publications are incorporated by reference in their entirety. In the event that there is a plurality of definitions for a term herein, those m this section prevail unless stated otherwise.VABIO.002WO2 PCT APPLICATION

[0069] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0070] The articles “a” and “an” are used herein to refer to one or to more than one (for example, at least one) of the grammatical object of the article, unless the context dictates otherwise. By way of example, “an element” means one element or more than one element.

[0071] The terms “first,” “second,” and “third” used in combination with substances are intended to designate distinguishable features of similar substances and do not imply any particular order unless otherwise specified.

[0072] As used herein, the term “including” as well as other forms such as “includes” and “included” is not limiting.

[0073] Throughout this specification, unless the context requires otherwise, the words “comprise,” “comprises,” and “comprising” will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements. By “consisting of” is meant including, and limited to, whatever follows the phrase “consisting of.” Thus, the phrase “consisting of’ indicates that the listed elements are required or mandatory, and that no other elements may be present. By “consisting essentially of,” it is meant including any elements listed after the phrase and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase “consisting essentially of’ indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present depending upon whether or not they materially affect the activity or action of the listed elements.

[0074] As used herein, the term “compound” has its usual meaning and thus can refer to a chemical or biological material. N on-limiting examples of a compound include an atom, small molecule, large molecule, peptide, protein, drug, vesicle, or any combination thereof.

[0075] As used herein, the term “composition” has its usual meaning and thus can refer to a chemical or biological compound or substance, or a mixture or combination of two or more such compounds or substances. In some embodiments, the composition comprises an atom, small molecule, large molecule, peptide, protein, salt, buffer, or any combination thereof. In some embodiments, the composition is formulated for a specific use, such as for administration to a subject.VABIO.002WO2 PCT APPLICATION

[0076] As used herein, “derivative” has its usual meaning and thus can refer to a chemical or biological material that is modified. In some embodiments, the derivative retains the activity of the parent material. For example, an estrogen derivative may comprise a modified estrogen that retains at least some of the function of estrogen.

[0077] As used herein, the terms “pharmaceutical composition” or “pharmaceutical formulation” may include a chemical or biological compound or substance, or a mixture or combination of two or more such compounds or substances, intended for use in the cure, treatment, prevention, or diagnosis of a disease or symptom.

[0078] As used herein, the terms “therapeutic agent” and “pharmaceutical agent” has their usual meaning and thus can refer to a compound or composition with a known therapeutic function. In some embodiments, this function is used in the cure, treatment, prevention, or diagnosis of a disease or symptom. In some embodiments, a therapeutic agent comprises a drug. In some embodiments, a therapeutic agent comprises a composition that includes a drug.

[0079] As used herein, “carrier” may include a substance that serves as a vehicle for improving the efficacy of delivery or the effectiveness of a pharmaceutical composition, or both.

[0080] As used herein, “pharmaceutically acceptable” when used to define a carrier, whether diluent or excipient, may include a substance that is compatible with other ingredients in a formulation and does not exert deleterious effects to the recipient thereof. As used herein, “pharmaceutically acceptable” has its plain and ordinary meaning as understood in light of the specification and may include carriers, excipients, and / or stabilizers that are nontoxic to the cell or mammal being exposed thereto at the dosages and concentrations employed or that have an acceptable level of toxicity. A “pharmaceutically acceptable” “diluent,” “excipient,” and / or “earner” as used herein have their plain and ordinary meaning as understood in light of the specification and may include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, that are compatible with administration to humans, cats, dogs, or other vertebrate hosts. Typically, a pharmaceutically acceptable diluent, excipient, and / or carrier is a diluent, excipient, and / or earner approved by a regulatory agency of a Federal, a state government, or other regulatory agency, or listed in the U. S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, including humans as well as non-human mammals, such as cats and dogs. The term diluent, excipient, and / or earner can refer to a diluent, adjuvant, excipient, or vehicle with which the pharmaceutical formulation isVABIO.002WO2 PCT APPLICATIONadministered. Such pharmaceutical diluent, excipient, and / or earners can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin. Water, saline solutions, and aqueous solutions of dextrose and glycerol can be used as liquid diluents, excipients, and / or carriers, particularly for injectable solutions. Suitable pharmaceutical diluents and / or excipients include sugars, starch, glucose, fructose, lactose, sucrose, maltose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, salts, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, and the like. A non-limiting example of a physiologically acceptable carrier is an aqueous pH-buffered solution. The physiologically acceptable carrier may also include one or more of the following: antioxidants, such as ascorbic acid, low molecular weight (less than about 10 residues) polypeptides, proteins, such as serum albumin, gelatin, immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidone, amino acids, carbohydrates such as glucose, mannose, or dextrins, chelating agents such as EDTA, sugar alcohols such as glycerol, erythritol, threitol, arabitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fucitol, iditol, inositol, isomalt, maltitol, or lactitol, salt-forming counterions such as sodium, and nonionic surfactants such as TWEEN®, polyethylene glycol (PEG), and PLUR0N1CS®. The formulation, if desired, can also contain minor amounts of wetting, bulking, emulsifying agents, or pH buffering agents. These formulations can take various forms, including solutions, suspensions, emulsions, and sustained-release formulations. The formulation should be suitable for the mode of administration.

[0081] The term “pharmaceutically acceptable salts” has its plain and ordinary meaning as understood in light of the specification and includes relatively non-toxic, inorganic and organic acid, or base addition salts of compositions or excipients, including, without limitation, analgesic agents, therapeutic agents, other materials, and the like. Examples of pharmaceutically acceptable salts include those derived from mineral acids, such as hydrochloric acid and sulfuric acid, as well as those derived from organic acids, including ethanesulfonic acid, benzenesulfonic acid, and p-toluenesulfomc acid, among others. Examples of suitable inorganic bases for the formation of salts include the hydroxides, carbonates, and bicarbonates of ammonia, sodium, lithium, potassium, calcium, magnesium, aluminum, zinc, and the like. Salts may also be formed with suitable organic bases, including those that are non-toxic and strong enough to form such salts. For example, the class of such organic bases may include but are not limited to mono-, di-, and trialkylamines, including methylamine, dimethylamine, and triethylamine; mono-, di-, orVABIO.002WO2 PCT APPLICATIONtrihydroxyalkylamines, including mono-, di-, and triethanolamine; amino acids, including glycine, arginine, and lysine; guanidine; N-methylglucosamine; N-methylglucamine; L-glutamine; N-methylpiperazine; morpholine; ethylenediamine; N-benzylphenethylamine; trihydroxymethyl aminoethane.

[0082] As used herein, “excipient” may include a pharmacologically inactive substance or compound that is formulated in combination with a pharmacologically active ingredient of a pharmaceutical composition, wherein an excipient may include a bulking agent, a filler, a diluent, or a product for drug solubility, drug absorption, or for modulating a pharmacokinetic property of an active ingredient of a pharmaceutical composition.

[0083] The terms “effective amount” or “effective dose” as used herein have their plain and ordinary meaning as understood in light of the specification, and may include an amount of a recited composition or compound that results in an observable effect. Actual dosage levels of active ingredients in an active composition of the presently disclosed subject matter can be varied so as to administer an amount of the active composition or compound that is effective to achieve the desired response for a particular subject and / or application. The selected dosage level will depend upon a variety of factors, including, but not limited to, the activity of the composition, formulation, route of administration, combination with other drugs or treatments, severity of the condition being treated, and the physical condition and prior medical history of the subject being treated. In some embodiments, a minimal dose is administered, and the dose is escalated in the absence of dose-limiting toxicity to a minimally effective amount. Determination and adjustment of an effective dose, as well as evaluation of when and how to make such adjustments, are contemplated herein.

[0084] The terms “function” and “functional” as used herein have their plain and ordinary meaning as understood in light of the specification, and may include a property exhibited by a molecule, compound, or composition, including, for example, a biological, enzymatic, or therapeutic activity.

[0085] “Regenerative activity'” or “regeneration” or “regenerative properties” (and grammatical equivalents thereof) as used herein has its customary and ordinary' meaning as understood by one of ordinary skill in the art in view of the present disclosure. “Regeneration” can refer to any process or quality that initiates, increases, modulates, promotes, supports, and / or directs the growth, regrowth, repair, functionality, patterning, connectivity, strengthening, vitality,!VABIO.002WO2 PCT APPLICATIONand / or the natural wound healing process of weak, damaged, lost, and / or degenerating tissue. These terms can also refer to any process or quality that initiates, increases, modulates, promotes, supports, and / or directs the growth, strengthening, functionality, vitality, toughness, potency, and / or health of weak, tired, and / or normal tissue. Regenerative activity can encompass the stimulation of angiogenesis, reduction of fibrosis, reduction of inflammation, differentiation, migration, and immunoregulation. Markers associated with regenerative activity can include heat shock proteins — such as calreticulin, HSP20, or HSP90 — stress-associated proteins — such as phosphatidylserine — or growth factor receptors — such as the IL-3 receptor, VEGF receptor, CXCL12, or c-Met. Markers associated with regenerative activity can also include tetraspanins such as CD9, CD63, or CD81, ALIX, ESCRT-III components such as CH1MP4B, ESCRT-I components such as TSG101, Rub GTPases, flotillin, heat shock proteins such as Hsp60, Hsp70, or Hsp90, growth factors such as PDGF-BB, FGF-2, VEGF-A, or I IGF, cytokines such as TGF-P, microRNAs such as miR-1246, miR-21, miR-140-5p, miR-92a-3p, and miR-100-5p, pluripotency-related markers such as PODXL, specific surface markers based on stem cell source, such as BMSC-exos ---which express markers such as CD63, CD9, and TSG101 -ADSC-exos — which express markers such as CD9, CD63, and CD81 - hfMSC-exos ---which express markers such as CD13, CD9, CD63, CD105, CD81, CD29, CD44, CD49e, and SSEA-4 -and iPSC-exos --- which express markers such as CD9, TSG101, and SSEA-1. Markers associated with regenerative activity can also include CAV1, FLOT1, FLOT2, PR0M1, CD63, CD9, or LAMP1.

[0086] The term “marker” has its standard scientific meaning and may include a biological molecule whose presence, concentration, activity, or phosphorylation state may be detected and used to identify the phenotype of a cell. Non-limiting examples of markers include heat shock proteins, stress-associated proteins, growth factor receptors, or a combination thereof.

[0087] The term “regenerative cell” as used herein has its customaiy and ordinary meaning as understood by one of ordinary skill in the art m view of the present disclosure. Regenerative cells encompass a diverse group of cells, and may include stem cells, committed progenitor cells, and differentiated cells.

[0088] The term “stem cell” has its standard scientific meaning and may include undifferentiated cells that can proliferate and have the capacity to both self-renew and differentiate to one or more types of specialized cells.VABIO.002WO2 PCT APPLICATION

[0089] The term “totipotent stem cell” has its standard scientific meaning and may include, for example, a stem cell capable of differentiating into any cell type m the body. In some embodiments, totipotent stem cells may include those of the germ line, as well as both embryonic and extraembryonic tissues, such as the placenta.

[0090] “Pluripotent stem cell” or “pluripotent cell” refers to a type of stem cell that has the ability, under appropriate conditions, to produce progeny of several different cell types that are derivatives of any one of the three germinal layers (endoderm, mesoderm, and ectoderm). Pluripotent stem (PS) cells are capable of forming teratomas. Examples of pluripotent stem cells are embryonic stem (ES) cells, embryonic germ stem (EG) cells, embryonal carcinoma stem (EC) cells, and induced pluripotent stem cells (1PS or IPSCs). Pluripotent stem cells may be from any one of a variety of organisms, including, e.g., human, primate, non-human primate, canine, feline, murine, equine, porcine, avian, camel, bovine, ovine, etc.

[0091] ‘Induced pluripotent stem cells” (IPSCs) describe a type of stem cell that is generated from adult (mature) cells, such as skin or blood cells, which have been reprogrammed back into an embryonic-like pluripotent state by means of chemicals, genes, reprogramming factors, micro ribonucleic acids (microRNA), small molecules, other factors / conditions, or a combination thereof. Induced pluripotent stem cells may be from any one of a variety of organisms, including, e.g., human, primate, non-human primate, canine, feline, murine, equine, porcine, avian, camel, bovine, ovine, etc.

[0092] “Multipotent stem cells” or “multipotent cells” or “multipotency” describes a stem cell's capacity to differentiate into multiple, but limited, cell types within a specific lineage or tissue type. Examples of multipotent stem cells include hematopoietic stem cells, mesenchymal stem cells (MSCs), neural stem cells, and epithelial stem cells. Multipotent stem cells may be from any one of a variety of organisms, including, e.g., human, primate, non-human primate, canine, feline, murine, equine, porcine, avian, camel, bovine, ovine, etc.

[0093] “Mesenchymal stem cells” (MSCs) are a type of multipotent stem cell with the ability to differentiate into various cell types. MSCs are typically a type of natural stem cell that can be isolated from various biological sources, including bone marrow (BM-MSCs), adipose tissue (AD-MSCs), umbilical cord (UC-MSCs), umbilical cord blood, dental pulp, placenta, synovial fluid, and others.VABIO.002WO2 PCT APPLICATION

[0094] “Mesenchymal stromal cells” refer to MSCs that are isolated from a sample, and may comprise a mixed population of stem-like and non-stem cells. In some embodiments, mesenchymal stromal cells can be isolated from bone marrow, adipose tissue, or the umbilical cord. In some embodiments, “mesenchymal stromal cells” refers to the bulk population, unless the sternness of these cells is proven.

[0095] “Human umbilical mesenchymal stem cells” (HUMSCs or huMSCs) or “human umbilical cord-derived MSCs” (hucMSCs) or “human umbilical cord mesenchymal stromal cells” (hucMSCs) are herein used interchangeably and refer to a type of multipotent, mesenchymal stem cell. In some embodiments, these cells are derived from the Wharton’s jelly of the umbilical cord. Wharton's jelly is a gelatinous substance found within the umbilical cord, primarily composed of mucopolysaccharides like hyaluronic acid and chondroitin sulfate, which contains mesenchymal stem cells (MSCs), often referred to as HUMSCs or hucMSCs.

[0096] “Hematopoietic stem cells” or “hematopoietic progenitor cells” refer to a type of multipotent stem cell that is committed to a hematopoietic lineage but capable of further hematopoietic differentiation. Hematopoietic stem cells can develop into all types of blood cells, including white blood cells, red blood cells, and platelets. Hematopoietic stem cells are primarily found in bone marrow, but can also be found in peripheral blood and umbilical cord blood.

[0097] The term “differentiation” has its standard scientific meaning and may include the process by which an unspecialized (“uncommitted”) or less specialized cell acquires the features of a specialized cell, such as, for example, a blood cell or a muscle cell. A differentiated or differentiation-induced cell is one that has taken on a more specialized (“committed”) position within the lineage of a cell. The term “committed”, when applied to the process of differentiation, may include a cell that has proceeded m the differentiation pathway to a point where, under normal circumstances, it will continue to differentiate into a specific cell type or subset of cell types, and cannot, under normal circumstances, differentiate into a different cell type or revert to a less differentiated cell type.

[0098] The term “stem cell derivatives” describes cells or substances derived from stem cells. Non-limiting examples of stem cell derivatives include differentiated cells, extracellular vesicles (EVs), secreted factors such as cytokines and growth factors, and culture media (CM).

[0099] “Extracellular vesicles” as used herein describe a diverse group of membrane¬ bound vesicles and can include small extracellular vesicles, exosomes, microvesicles, apoptoticVABIO.002WO2 PCT APPLICATIONbodies, free mitochondria, ectosomes, oncosomes, and vesicular structures. “Small extracellular vesicles” (sEVs) are typically less than 200 nm in size. “Exosomes” are a subset of sEVs, are typically about 30-200 nm in size, and are formed inside endosomal compartments. “Microvesicles” (MVs) are typically about 100- 1000 nm in size and originate by budding directly from the plasma membrane. “Apoptotic bodies” are typically about 1-5 m and are released during programmed cell death.

[0100] “Human Umbilical Cord Mesenchymal Stem Celis-derived Exosomes” or “ucMSC-Ex” or “hucMSC-Exo” are herein used interchangeably and refer to exosomes derived from human umbilical cord mesenchymal stem cells,

[0101] The term “tetraspanin” refers to a protein family that organizes membrane microdomains, termed tetraspanin-enriched microdomains (TEMs) by forming clusters and interacting with a variety of transmembrane and cytosolic signaling proteins (see e.g., Hemler et al. (2005) Nat, Rev. Mol. Cell. Biol, 6:801-811). Since tetraspanins are expressed on various types of endocytic membranes, they have been used in the art as exosomal markers. In some embodiments, the tetraspanin molecule may be selected from, but not limited to TSPANI, TSPAN2, TSPAN3, TSPAN4 (also known as NAG-2), TSPAN5, TSPAN6, TSPAN7 (also known as CD231, TALLA-1, A15), TSPAN8 (also known as CO-029), TSPAN9 (also known as NET-S1, TSPAN10 (also known as OCULOSPANIN), TSPAN11 (also known as CD151-like), TSPAN12 (also known as NET-2), TSPAN13 (also known as NET-6), TSPAN14, TSPAN15 (also known as NET-7), TSPAN16 (also known as TM4-B), TSPAN17, TSPAN18, TSPAN19, TSPAN20 (also known as UPlb, UPK1B), TSPAN21 (also known as UPla, UPK1A), TSPAN22 (also known as RDS, PRPH2), TSPAN23 (also known as R0M1), TSPAN24 (also known as CD151), TSPAN25 (also known as CD53), TSPAN26 (also known as CD37), TSPAN27 (also known as CD82, KAI1), TSPAN28 (also known as CD81, TAPA-1), TSPAN29 (also known as CD9), TSPAN30 (also known as CD63), TSPAN31 (also known as SAS), TSPAN32 (also known as TSSC6), TSPAN33, or a combination thereof.

[0102] The term “bioactive lipids” has its standard scientific meaning and may include molecules that respond to specific stimuli, affecting cell function by signaling and regulating cell metabolism. Examples of bioactive lipids that can be released by extracellular vesicles include eicosanoids, cholesterol, lysophosphatidylcholine, and arachidonic acid.VABIO.002WO2 PCT APPLICATION

[0103] The term “angiogenesis” has its standard scientific meaning and may include a biological process that involves the sprouting of new blood vessels from pre-existing ones and plays a crucial role in disease development and progression.

[0104] The terms “toll-like receptor(s)” or “toll-like receptor” or “TLR(s)” has their standard scientific meaning and may include a class of pattern recognition receptors that play an essential role in innate immunity, by recognizing invasion of microbial pathogens and initiating intracellular signal transduction pathways to trigger expression of genes, the products of which can control innate immune responses. Examples of TLRs include TLR4 and TLR3,

[0105] The term “fibrosis” has its standard scientific meaning and may include the formation of excessive connective tissue in an organ or tissue, commonly in response to damage or injury.

[0106] “Heat shock proteins” (HSP(s) or Hsp or hsp), also known as stress proteins, are a family of proteins produced by cells in response to stressful conditions, such as heat. Examples of HSP families include Hsp20, Hsp60, Hsp70, Hsp90, Hsp110, small HSPs, and large HSPs.

[0107] The term “growth factor receptor” as used herein is a protein on the surface of a cell that binds to a growth factor, initiating a signaling cascade that regulates cell growth, proliferation, differentiation, and survival. Growth factor receptors are typically transmembrane proteins with extracellular, transmembrane, and cytoplasmic domains. Examples of growth factor receptors include IL-3 receptor, VEGF receptor, CXCL12, c-met, fibroblast growth factor (bFGF), and epidermal growth factor (EGF).

[0108] The term “microRNA” or “micro-RNA” or “miRNAs” as used herein has its customary and ordinary meaning as understood by one of ordinary skill in the art in view of the present disclosure. MicroRNAs are a class of non-coding RNAs that often play important roles in regulating gene expression.

[0109] The term “messenger RN A” or “mRN A” as used herein has its customary and ordinaiy meaning as understood by one of ordinary’ skill in the art in view of the present disclosure. mRNA refers to a polynucleotide that encodes at least one polypeptide.

[0110] The term “antigen” as used herein has its customary and ordinaiy meaning as understood by one of ordinary skill in the art in view’ of the present disclosure. As used herein, an “antigen” refers to any substrate that can be specifically bound by an antibody molecule. ExamplesVABIO.002WO2 PCT APPLICATIONof antigens include: stage-specific embryonic antigens such as SSEA-3 and SSEA 4, Tra-1-60, Tra-1-81, Oct-3 / 4, Cripto, gastrin-releasing peptide (GRP) receptor, podocalyxin-like protein (PODXL), Rex-1, GCTM-2, Nanog, human telomerase reverse transcriptase (hTERT), CD9, CD34, c-kit, OCT-4, Nanog, CXCR-4, CD13, CD29, CD44, CD166, CD90, CD105, SH-3, SH-4, TRA-1-60, TRA-1-81, SSEA-4, Sox-2, flk-1, Stro-1, CD73, CD31, CD146, vascular endothelial- cadherin, CD133, CXCR-4, Tra-2-54, HLA class I, CD49b, CD105, DAZL, Runx-1, CD45, HLA Class II, RC-2, 3CB2, BLB, Sox-2, GLAST, Pax 6, nestin, Muashi-1, NCAM, A2B5, and prominin.

[0111] “Autologous” materials as used herein are those taken from a patient’s own tissues, cells, or DNA. “Allogeneic” materials as used herein are genetically dissimilar from the patient, although both the donor and the patient are individuals of the same species. “Xenogeneic” materials, as used herein, are obtained from a species different from that of the patient.

[0112] “Express” or “expression” as used herein refers to the presence of a gene product (e.g., mRNA and / or protein) in or associated with a cell or a group of cells at a level above a relevant reference level (e.g., a predetermined level for detecting the gene product, or a level of the gene product that effectively differentiates between populations of cells known to either express or not express the gene product). In some cases, the reference level is a background level of expression.

[0113] The term “culture” or “cell culture” refers to the maintenance, growth, and / or differentiation of cells in an in vitro environment.

[0114] “Cell culture media,” “culture media” (CM) (singular “medium” in each case), “supplement,” and “media supplement” refer to nutritive compositions that cultivate cell cultures. The term “culture media” is sometimes used interchangeably with “conditioned media” to describe culture media that have been previously exposed to and partially consumed by cells, containing secreted cellular products.

[0115] The term “cultivates” or “maintains” refers to the sustaining, propagating (growing), and / or differentiating of cells outside of tissue or the body, for example, in a sterile plastic (or coated plastic) cell culture dish or flask. “Cultivation,” or “maintaining,” may utilize a culture medium as a source of nutrients, hormones, and / or other factors helpful to propagate and / or sustain the cells.VABIO.002WO2 PCT APPLICATION

[0116] As used herein, the term "therapeutic target" has its plain and ordinary meaning as understood in light of the specification and may include a gene or gene product that, upon modulation of its activity (e.g., by modulation of expression, biological activity, and the like), can provide for modulation of the disease phenotype. As used throughout, "modulation" refers to an increase or decrease in the indicated phenomenon (e.g., modulation of a biological activity refers to either an increase or a decrease in a biological activity).

[0117] “Effective amount” as used herein refers to an amount sufficient to bring about a desired result. “Therapeutically effective amount” as used herein refers to an amount sufficient for a therapeutic agent to bring about a therapeutically desired result.

[0118] The term “administering” includes contact with the skin, tissue, mucus, or fluid of a subject. It includes topical contact, administration as a suppository,, intravaginal, intravenous, intraperitoneal, intramuscular, intralesional, intratumoral, intrathecal, intranasal, or subcutaneous administration, or the implantation of a slow-release device, such as a mini-osmotic pump, to a subject. Administration can be by any route unless specifically stated, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intra-tumoral, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, and transdermal patches. By “co-administer,” it is meant that a first compound described herein is administered at the same time, just prior to, or just after the administration of a second compound described herein. The preferred administration of the present disclosure comprises contact with a subject’s reproductive glands or tissues, such as through transvaginal delivery and / or intravaginal delivery.

[0119] As used herein, “administration” (or other forms such as “administering”) may include an act of providing a substance or compound to a subject in need thereof.

[0120] As used herein, the terms "treatment" or "treat" (and grammatical equivalents thereof) refer to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of patient at risk of contracting the disease or suspected to have contracted the disease as well as patients who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse. The treatment may be administered to a patient having a medical disorder or who ultimately may acquire the disorder, inVABIO.002WO2 PCT APPLICATIONorder to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a patient beyond that expected in the absence of such treatment. The terms also include: (a) preventing the disease or symptom from occurring in a subject which may be predisposed to the disease or symptom but has not yet been diagnosed as having it; (b) inhibiting the disease symptom, e.g., arresting or delaying its development; and / or (c) relieving the disease symptom, e.g., causing regression of the disease or symptom.

[0121] The term “active ingredient” as used herein is to be given its ordinary and customary meaning to a person of ordinary skill in the art (and is not to be limited to a special or customized meaning) and may include a component that confers a function. In some embodiments, the component has a direct or indirect effect on the cure, mitigation, treatment, diagnosis, or prevention of a disease or condition in the human or animal body.

[0122] “Preventing” in reference to a disease, disorder, or condition refers to preventing a disease, disorder, or condition, e.g,, causing the clinical symptoms of the disease, disorder, or condition not to develop. As used herein, the term “prevent,” “prevents,” or “prevention” as well as “inhibit,” “inhibits,” or “inhibiting” (and grammatical equivalents thereof) may also refer to a delay in the onset of a disease or disorder or the lessening of symptoms upon onset of the disease or disorder. The terms are not meant to imply complete abolition of disease and encompass any type of prophylactic treatment that reduces the incidence of the condition or delays the onset and / or progression of the condition.

[0123] As used herein, the term “isolated” has its plain and ordinary meaning as understood in light of the specification, and may include to a substance and / or entity that has been (1) separated from at least some of the components with which it was associated when initially produced (whether in nature and / or in an experimental setting), and / or (2) produced, prepared, and / or manufactured by the hand of man. Isolated substances and / or entities may be separated from equal to, about, at least, at least about, not more than, or not more than about, 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 98%, about 99%, substantially 100%, or 100% of the other components with which they were initially associated (or ranges including and / or spanning the aforementioned values). In some embodiments, isolated agents are, are about, are at least, are at least about, are not more than, or are not more than about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, aboutVABIO.002WO2 PCT APPLICATION94%, about 95%, about 96%, about 97%, about 98%, about 99%, substantially 100%, or 100% pure (or ranges including and / or spanning the aforementioned values). As used herein, a substance that is “isolated” may be “pure” (e.g., substantially free of other components). As used herein, the term “isolated cell” may refer to a cell not contained in a multicellular organism or tissue. The term includes a cell that is removed from some or all components as it is found in its natural environment, for example, isolated from a tissue or biopsy sample. The term also includes a cell that is removed from at least one, some, or all components, as the cell is found in non-naturally occurring environments, for example, isolated from a cell culture or cell suspension. Therefore, an isolated cell is partly or completely separated from at least one component, including other substances, cells, or cell populations, as it is found in nature or as it is grown, stored, or subsisted in non-naturally occurring environments.

[0124] The term “purity” of any given substance, compound, or material, as used herein, has its plain and ordinary meaning as understood in light of the specification and may include the actual abundance of the substance, compound, or material relative to the expected abundance. For example, the substance, compound, or material may be at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% pure, including all decimals in between. Purity may be affected by unwanted impurities, including but not limited to nucleic acids, DNA, RNA, nucleotides, proteins, polypeptides, peptides, amino acids, lipids, cell membrane, cell debris, small molecules, degradation products, solvent, carrier, vehicle, or contaminants, or any combination thereof. In some embodiments, the substance, compound, or material is substantially free of host cell proteins, host cell nucleic acids, plasmid DNA, contaminating viruses, proteasomes, host cell culture components, process-related components, mycoplasma, pyrogens, bacterial endotoxins, and adventitious agents. Purity can be measured using technologies including but not limited to electrophoresis, SDS-PAGE, capillary electrophoresis, PCR, rtPCR, qPCR, chromatography, liquid chromatography, gas chromatography, thin layer chromatography, enzyme-linked immunosorbent assay (ELISA), spectroscopy, UV-visible spectrometry, infrared spectrometry, mass spectrometry, nuclear magnetic resonance, gravimetry, or titration, or any combination thereof.

[0125] The term “ex vivo” generally refers to activities that take place outside an organism, such as experimentation or measurements performed on living tissue in an artificial environment outside the organism.VABIO.002WO2 PCT APPLICATION

[0126] The term “young animal” as used herein refers to a non-human animal that has not yet reached reproductive maturity or full physical development. The age or developmental stage constituting a “young animal” may vary by species. For example, in mammals, a young animal may include neonates, juveniles, or pre- weaning animals.

[0127] “Scaffold” as used herein is a broad term and is to be given its ordinary meaning to a person of ordinary’ skill in the art (and is not to be limited to a special or customized meaning), and refers without limitation to porous materials or structures which is suitable for structural support and / or providing an environment conducive to cell activity, such as a material or structure which mimics an extracellular matrix in biological tissue or organ systems. “Scaffolds” may also provide support for cells to adhere, migrate, and form functional tissues. “Scaffold” may include a material or structure that aids in support and / or interaction of other molecules, compositions, drugs cells, tissues, systems, or any combination thereof. In some embodiments, a scaffold comprises a porous material and / or a porous structure. In some embodiments, the scaffold is suitable for structural support and / or providing an environment conducive to cell activity, A non-limiting example of a scaffold includes a material or structure that mimics an extracellular matrix in biological tissue or organ systems.

[0128] The terms “individual”, “subject”, or “patient” as used herein have their plain and ordinary meaning as understood in light of the specification, and may include a human or a non-human mammal, e.g., a dog, a cat, a mouse, a rat, a cow, a sheep, a pig, a goat, a non-human primate, or a bird, e.g., a chicken, as well as any other vertebrate or invertebrate. The term “mammal” is used in its usual biological sense. Thus, it includes, but is not limited to, primates (including simians such as chimpanzees, apes, and monkeys), as well as humans, cattle, horses, sheep, goats, swine, rabbits, dogs, cats, rodents (including rats and mice), guinea pigs, and the like. “Woman” and “female” as used herein are used interchangeably and are defined to mean an individual comprising some or all portions of the female genitourinary tract. A “woman” is not limited by age, and will be understood to include prepubescent, pubescent, pre-menopausal, and post-menopausal individuals. A “woman” or “female” is not necessarily limited to humans.

[0129] The terms “postpartum,” “childbirth,” and “vaginal delivery’” as used herein are not limited to the delivery’ of live offspring.

[0130] The terms “female genitourinary system” or “female genitourinary’ tract” has their standard scientific meaning and may include systems such as the female genitals,VABIO.002WO2 PCT APPLICATIONreproductive organs, and urinary organs. Non-limiting examples of female reproductive organs include the ovaries, fallopian tubes, uterus, cervix, vagina, and vulva. Non-limiting examples of urinary organs include the kidneys, ureters, bladder, and urethra.

[0131] “Menopause” has its standard scientific meaning and may include the period in a female mammal’s life when her menstrual periods stop permanently, typically due to decreased hormone production and loss of follicular activity by the ovaries. “Ovariectomy” or “oophorectomy” as used herein refers to the removal of at least one ovary — typically two ovaries — and can also induce menopause.

[0132] The term “menopausal” has its standard scientific meaning and may include the period of time leading up to and during menopause.

[0133] The term “postmenopausal” as used herein refers to the period of time following the substantial cessation of menopause caused either by natural means or surgical inducement,

[0134] “Genitourinary syndrome of menopause” (GSM) as used herein has its customary and ordinary meaning as understood by one of ordinary skill in the art in view of the present disclosure. GSM describes the multiple symptoms resulting from estrogen deficiency in the female genitourinary tract, including the vagina, labia, urethra, and bladder. This syndrome includes genital symptoms of dryness, burning, and irritation; urinary symptoms of dysuria, urgency, and recurrent urinary tract infections (UTIs); and sexual symptoms of pain and dryness. Vulvovaginal atrophy is a component of GSM. GSM can affect women of any age and typically develops in the setting of hypoestrogenism associated with natural menopause; however, hypoestrogenic states may occur in other settings.

[0135] “Postpartum vaginal damage” (PVD) as used herein has its customary and ordinary meaning as understood by one of ordinary skill in the art in view of the present disclosure. PVD commonly causes tears to the vagina and the surrounding perineum, caused by stretching during childbirth. There are four main degrees of postpartum vaginal damage. First-degree tears are the least severe and involve tears to the perineum and urethra. Second-degree tears are more severe, as they may go deeper into the vagina and typically require stitches. Second-degree tears are the common form of PVT). Third-degree tears extend from the vagina to the anus and damage the anal sphincter muscles. Finally, fourth-degree tears are the least common and the most severe, and extend from the vagina, through the perineal area and anal sphincter muscles, into the rectum.VABIO.002WO2 PCT APPLICATION

[0136] “Suppository” or “vaginal suppository” as used herein refers to a solid or semi¬ solid medical preparation designed to be inserted into the vagina to dissolve. The terms “suppository'” or “vaginal suppository'” as used herein also describe suppository-type devices.

[0137] “Biocompatible” as used herein is a broad term and is to be given its ordinary meaning to a person of ordinary skill in the art, and may include a material’s ability to perform an appropriate host response in a specific situation. In some embodiments, biocompatibility refers to a material’s interaction with a tissue or other part of a living body or system without causing undesirable or harmful effects, such as inflammation or irritation. In some embodiments, biocompatibility refers to the ability of a biomaterial to perform its desired function with respect to a medical therapy, without eliciting any undesirable local or systemic effects in the recipient or beneficiary of that therapy, but generating the most appropriate beneficial cellular or tissue response in that specific situation, and optimizing the clinically relevant performance of that therapy. In some embodiments, biocompatibility refers to the ability of a biomaterial to perform its desired function without causing a negative reaction with the surrounding tissue, or with the subject’s immune system.

[0138] A “biomaterial” as used herein is a broad term and is to be given its ordinary meaning to a person of ordinary skill in the art and may include a substance that has been engineered to interact with a biological molecule, cell, tissue, system, or individual. In some embodiments, a biomaterial is a biological protein, such as an enzyme. In some embodiments, the biomaterial has been engineered for medical purposes, such as treating, augmenting, repairing, or replacing tissue function in the body. In some embodiments, the biomaterial is naturally occurring. In some embodiments, the biomaterial is synthetic or is a modified variant of a naturally occurring material

[0139] “Excipient” as used herein is a broad term and is to be given its ordinary meaning to a person of ordinary skill in the art, and may include to a substance that is added to a pharmaceutical composition to provide, without limitation, bulk, consistency, stability, binding ability, lubrication, disintegrating ability, etc., to the composition. A “diluent” is a non-limiting example of a type of excipient. In some embodiments, an excipient refers to a pharmacologically inactive substance or compound that is formulated in combination with a pharmacologically active ingredient of a pharmaceutical composition, wherein an excipient may include a bulking agent, aVABIO.002WO2 PCT APPLICATIONfiller, a diluent, or a product for drug solubility, drug absorption, or for modulating a pharmacokinetic property of an active ingredient of a pharmaceutical composition.

[0140] “Pharmaceutically acceptable” as used herein is a broad term, and is to be given its ordinary’ meaning to a person of ordinary skill in the art (and is not to be limited to a special or customized meaning), and may include compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for contact with tissues of human beings and animals without excessive toxicity’, irritation, allergic response, or other problem or complications commensurate with a reasonable risk / benefit ratio.

[0141] The terms “about,” “approximately,” or “roughly” mean within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will 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, per the practice in the art. Alternatively, “about” can mean a range of up to 20%, up to 10%, up to 5%, and 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, and within 2-fold of a value. Where particular values are described in the application and claims, unless otherwise stated, the term “about” means within an acceptable error range for the particular value. The terms “about”, “approximately”, or “roughly”, as used herein, for example, define the values and ranges of weights, volumes, or lengths, meaning that the indicated values and / or range limits can vary within ±20%, e.g., within ±10%, including within ±5%. The use of “about” before a number includes the number itself. For example, “about 5” provides express support for “5.”

[0142] The term “size” as used herein to describe biomolecules, particles, cellular components, or secreted cellular components such as extracellular vesicles (herein “specimen(s)”), tends to describe the approximate diameter of the specimen, but does not limit qualifying specimens to those that are spherically or cylindrically-shaped. Therefore, “size” can be used to describe irregularly-shaped specimens. Similarly, the term “diameter” encompasses irregular¬ shaped specimens and is not limited to spherically or cylindrically shaped specimens. Diameter and size of specimens should be determined by the methods of one of ordinary skill in the art in view of the present disclosure.

[0143] The term “chitosan” as used herein is given its usual meaning and may include a linear polysaccharide comprising at least one D-glucosamme (deacetylated unit) and / or at leastVABIO.002WO2 PCT APPLICATIONone A-acetyl-D-glucosamine (acetylated unit). In some embodiments, the chitosan comprises at least one P-(l-~>4)-linked D-ghicosamine. In some embodiments, chitosan may be formed through the deacetylation of chitin, such as through contacting chitin with aqueous sodium hydroxide. In some embodiments, chitosan is a natural, biodegradable, and cationic polymer with use in the field of tissu e engineering. In some embodiments, chitosan is modified. In some embodiments, chitosan is phosphorylated, thiolated, quaternized, or any combination thereof.

[0144] The term “collagen” as used herein is given its usual meaning and may include protein collagen. In some embodiments, the collagen is from human and / or mammalian sources. In some embodiments, the collagen comprises at least one of types I-XXVIII, In some embodiments, the collagen comprises at least one of types I, II, III, IV, or V, In some embodi ents, collagen is modified. In some embodiments, collagen is in its natural protein form present in at least one of: bones, joints, muscles, skin, or any combination thereof. In some embodiments, collagen is in its monomer form. In some embodiments, collagen is a triple helix. In some embodiments, collagen is used as part of tissue regeneration in order to increase the proliferation of new cells.

[0145] The term “Carboxymethyl cellulose (CMC)” as used herein is a water-soluble derivative of cellulose, which is widely considered to be biocompatible and is commonly used in a range of biomedical and pharmaceutical applications. CMC exhibits characteristics such as nontoxicity, biodegradability, good hydrophilicity, low immunogenicity, and versatility in formulation, making it suitable as a cross-linking material.Stem Cells

[0146] A stem cell can refer to any undifferentiated cell, so as those that can proliferate and have the capacity to both self-renew and differentiate to one or more types of specialized cells. Nonlimiting examples of stem cells include hematopoietic stem cells, mesenchymal stem cells, mesenchymal stromal cells (MSCs), embryonic stem cells, umbilical cord stem cells, umbilical cord blood stem cells, placental stem cells, bone marrow stem cells, amniotic fluid stem cells, neuronal stem cells, circulating peripheral blood stem cells, germinal stem cells, adipose tissue derived stem cells, exfoliated teeth derived stem cells, hair follicle stem cells, dermal stem cells, parthenogenically derived stem cells, reprogrammed stem cells, and side population stem cells.VABIO.002WO2 PCT APPLICATIONInduced pluripotent stem cells (iPSCs)

[0147] In some embodiments, the regenerative cell population is a population of pluripotent or multipotent stem cells. In some embodiments, the pluripotent cells are induced pluripotent stem cells, somatic cell nuclear transfer-derived pluripotent stem cells, or parthenogenesis-derived stem cells. In some embodiments, stem cells are derived from an allogenic donor. In some embodiments, induced pluripotent stem cells (iPSCs) are used for treatment, and the cells were derived from the same patient as the one being treated.Hematopoietic Stem Cells

[0148] In some embodiments, the hematopoietic stem cells may express markers such as CD 133, CD34, CD45, c-kit, IL-3 receptor, c-mpl, c-met, stem cell factor receptor, or LAP. In some embodiments, the hematopoietic stem cells are capable of regenerating hematopoiesis in an immunodeficient mouse.Mesenchymal Stromal Cells (MSCs)

[0149] In some embodiments, the MSCs are preconditioned to stimulate enhanced production and efficacy of therapeutic extracellular vesicles. In some embodiments, this preconditioning is achieved by treating cells with hormones (including estrogen, progesterone, Follicle-Stimulating Hormone (FSH), Luteinizing Hormone (LH), Human Chorionic Gonadotropin (hCG), androgens, etc.). In some embodiments, this preconditioning is achieved by treating cells under conditions that induce cellular stress. This cellular stress can include nuclear translocation of NF-kappa B, HIF-1 alpha, heme oxygenase- 1, or TyrRS by more than 10% as compared to baseline. Cellular stress can also comprise hypoxic stress, hypotonic stress, hypertonic stress, radiation-induced stress, ultrasound-induced stress, and inflammation-induced stress (interferon gamma (IFN-y), interleukin (IL)-la / p, IL-25, IL-6, TNF-a, and IL-17, among others). In some embodiments, inflammation-induced stress results from the activation of one or more toll-like receptors. In certain embodiments, a toll like receptor is TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, or TLR10. In certain embodiments, a toll-like receptor is TLR4 or TLR3. A TLR4 receptor activator may comprise beta-glucan, low molecular weight hyaluronic acid, or histone-DNA complexes. The histone-DNA complexes may be derived from neutrophil extracellular traps. A TLR3 receptor activator may comprise Poly IC.VABIO.002WO2 PCT APPLICATION

[0150] In some embodiments, MSCs express one or more of the following markers: STRO-1, CD 105, CD54, CD 106, HLA-I markers, vimentin, ASMA, collagen- 1, fibronectin, LFA-3, ICAM-1, PECAM-1, P-selectm, L-selectin, CD49b / CD29, CD49c / CD29, CD49d / CD29, CD61, CD18, CD29, thrombomodulin, telomerase, CD10, CD13, STRO-2, VCAM-1, CD146, and THY- 1. In some embodiments, MSCs do not express substantial levels of HLA-DR, CD117, and CD45. In some embodiments, MSCs are characterized by the positive expression of surface markers CD73, CD90, and CD105. In some embodiments, MSCs are characterized by the absence of CD34 and CD45 surface markers, which are markers commonly associated with hematopoietic cells and thus help distinguish MSCs from hematopoietic cells.

[0151] In some embodiments, MSCs are derived from a group comprising bone marrow, adipose tissue, umbilical cord or umbilical cord blood, placental tissue, peripheral blood mononuclear cells, differentiated embryonic stem cells, and differentiated progenitor cells. In some embodiments, MSCs can be isolated from various sources, including bone marrow (BM-MSCs), adipose tissue (AD-MSCs), umbilical cord (UC-MSCs), umbilical cord blood, dental pulp, placenta, and synovial fluid, among others.Embryonic Stem Cells

[0152] In some embodiments, embryonic stem cells are totipotent. In some embodiments, the embryonic stem cells express at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 of the following antigens: stage-specific embryonic antigens (SSEA) 3, SSEA 4, Tra-1-60 and Tra-1-81, Oct-3 / 4, Cripto, gastrin-releasing peptide (GRP) receptor, podocalyxm-like protein (PODXL), Rex-1, GCTM-2, Nanog, human telomerase reverse transcriptase (hTERT), or any combination thereof.Umbilical Cord Stem Cells

[0153] In some embodiments, umbilical cord stem cells are derived from Wharton’s jelly (the gelatinous tissue of the cord). In some embodiments, umbilical cord stem cells are rich in mesenchymal stem / stromal cells (MSCs), which can differentiate into bone, cartilage, fat, and support tissue repair. In some embodiments, the umbilical cord stem cells are identified based on the expression of one or more antigens selected from the group comprising CD73, CD90 (Thy-1), CD 105 (Endoglin), CD44, CD29, and CD 166.VABIO.002WO2 PCT APPLICATIONUmbilical Cord Blood Stem Cells

[0154] In some embodiments, umbilical cord blood stem cells are multipotent. In some embodiments, umbilical cord blood stem cells are capable of differentiating into endothelial cells, smooth muscle cells, neuronal cells, or any combination thereof. In some embodiments, the umbilical cord blood stem cells are identified based on expression of at least one of the following antigens: SSEA-3, SSEA-4, CD9, CD34, c-kit, OCT-4, Nanog, and CXCR-4. In some embodiments, the umbilical cord blood stem cells do not express one or more markers selected from a group consisting of: CD3, CD34, CD45, and CD1 lb.Placental Stem Cells

[0155] In some embodiments, placental stem cells are isolated from the placental structure. In some embodiments, placental stem cells are identified based on the expression of one or more antigens selected from the group comprising Oct-4, Rex-1, CD9, CD13, CD29, CD44, CD166, CD90, CD105, SH-3, SH-4, TRA-1-60, TRA-1-81, SSEA-4, Sox-2, or any combination thereof.Bone Marrow Stem Cells

[0156] In some embodiments, bone marrow stem cells comprise bone marrow mononuclear cells. In some embodiments, said bone marrow stem cells are selected based on the ability to differentiate into one or more of the following cell types: endothelial cells, smooth muscle cells, or neuronal cells. In some embodiments, bone marrow stem cells are selected based on the expression of one or more of the following antigens: CD34, c-kit, flk-1, Stro-1, CD105, CD73, CD31, CD 146, vascular endothelial cadherin, CD 133, CXCR4, or any combination thereof. In some embodiments, bone marrow stem cells are enriched for expression of CD 133.Amniotic Fluid Stem Cells

[0157] In some embodiments, amniotic fluid stem cells are isolated by introducing a fluid extraction device into the amniotic cavity under ultrasound guidance. In some embodiments, amniotic fluid stem cells are selected based on expression of one or more of the following antigens: SSEA3, SSEA4, Tra-1-60, Tra-1-81, Tra-2-54, HLA class I, CD13, CD44, CD49b, CD105, Oct-4, Rex-1, DAZE, Runx-1, or any combination thereof. In some embodiments, amniotic fluid stemVABIO.002WO2 PCT APPLICATIONcells are selected based on the lack of expression of one or more of the following antigens: CD34, CD45, HLA Class II, or any combination thereof.Neuronal Stem Cells

[0158] In some embodiments, neuronal stem cells are selected based on the expression of one or more of the following antigens: RC-2, 3CB2, BLB, Sox-2, GLAST, Pax6, nestin, Muash-1, NCAM, A2B5, prominin, or any combination thereof.Circulating Peripheral Blood Stem Cells

[0159] In some embodiments, circulating peripheral blood stem cells are characterized by the ability to proliferate in vitro for a period of over 3 months. In some embodiments, circulating peripheral blood stem cells are characterized by expression of at least one of: CD34, CXCR4, CD117, CD113, c-met, or any combination thereof. In some embodiments, circulating peripheral blood stem cells lack substantial expression of an at least one differentiation-associated marker. In some embodiments, the at least one differentiation associated marker is selected from the group consisting of: CD2, CD3, CD4, CD11, CD1 la, Mac-1, CD14, CD16, CD19, CD24, CD33, CD36, CD38, CD45, CD56, CD64, CD68, CD86, CD66b, and I ILA-DR.Germinal Stem Cells

[0160] In some embodiments, germinal stem cells express an at least one marker selected from a group comprising: Oct4, Nanog, Dppa5, Rbm, cyclin A2, Text 8, Stra8, Dazl, betal- and alphab-integrins, Vasa, Fragilis, Nobox, c-Kit, Sca-1, and Rexl.Adipose Tissue-Derived Stem Cells

[0161] In some embodiments, adipose tissue-derived stem cells express an at least one marker selected from a group comprising: CD13, CD29, CD44, CD63, CD73, CD90, CD166, Aldehyde dehydrogenase (ALDH), and ABCG2. In some embodiments, adipose tissue-derived stem cells are a population of purified mononuclear cells extracted from adipose tissue that are capable of proliferating in culture for more than one month.Exfoliated Teeth Derived Stem CellsVABIO.002WO2 PCT APPLICATION

[0001] In some embodiments, exfoliated teeth-derived stem cells express an at least one marker selected from a group comprising: STRO-1, CD146 (MUC18), alkaline phosphatase, MEPE, and bFGF.Hair Follicle Stem Cells

[0002] In some embodiments, hair follicle stem cells express an at least one marker selected from a group comprising: cytokeratin 15, Nanog, and Oct-4. In some embodiments, hair follicle stem cells are capable of proliferating in culture for at least one month. In some embodiments, hair follicle stem cells secrete one or more of the foll owing proteins when cultured in: basic fibroblast growth factor (bFGF), endothelin-1 (ET-1), and stem cell factor (SCF).Dermal Stem Cells

[0162] In some embodiments, dermal stem cells express at least one marker selected from a group comprising: CD44, CD13, CD29, CD90, and GDI 05, In some embodiments, dermal stem cells are capable of proliferating in culture for at least one month.Parthenogenically Derived Stem Cells

[0163] In some embodiments, parthenogenically derived stem cells are generated by adding a calcium flux-inducing agent to activate an oocyte, followed by enrichment of cells expressing an at least one marker selected from the group including: SSEA-4, TRA-1-60, and TRA-1-81.Reprogrammed Stem Cells

[0164] In some embodiments, reprogrammed stem cells are selected from a group comprising: cells subsequent to a nuclear transfer, cells subsequent to a cytoplasmic transfer, cells treated with a DNA methyltransferase inhibitor, cells treated with a histone deacetylase inhibitor, cells treated with a GSK-3 inhibitor, cells induced to dedifferentiate by alteration of extracellular conditions, and cells treated with various combination of the mentioned treatment conditions. In some embodiments, the nuclear transfer involves introducing nuclear material into a substantially enucleated cell, where the nuclear material originates from a host ’hose genetic profile is intended to be dedifferentiated. In some embodiments, the cytoplasmic transfer involves introducing theVABIO.002WO2 PCT APPLICATIONcytoplasm of a cell with a dedifferentiated phenotype into a cell with a differentiated phenotype, thereby causing the cell with a differentiated phenotype to substantially revert to a dedifferentiated phenotype. In some embodiments, the DNA demethylating agent is selected from the group comprising 5 -azacytidine, psammaplin A, and zebularine. In some embodiments, the histone deacetylase inhibitor is selected from the group comprising valproic acid, trichostatin A, trapoxin A, and depsipeptide.Side Population Stem Cells

[0165] In some embodiments, the side population cells are identified based on the expression of the multidrug resistance transport protein (ABCG2) or their ability to efflux intracellular dyes, such as rhodamine- 123 and / or Hoechst 33342, In some embodiments, said cells are derived from tissue. Nonlimiting examples of tissue include pancreatic tissue, liver tissue, smooth muscle tissue, striated muscle tissue, cardiac muscle tissue, bone tissue, bone marrow tissue, bone spongy tissue, cartilage tissue, liver tissue, pancreas tissue, pancreatic ductal tissue, spleen tissue, thymus tissue, Peyer's patch tissue, lymph nodes tissue, thyroid tissue, epidermis tissue, dermis tissue, subcutaneous tissue, heart tissue, lung tissue, vascular tissue, endothelial tissue, blood cells, bladder tissue, kidney tissue, digestive tract tissue, esophagus tissue, stomach tissue, small intestine tissue, large intestine tissue, adipose tissue, uterus tissue, eye tissue, lung tissue, testicular tissue, ovarian tissue, prostate tissue, connective tissue, endocrine tissue, and mesentery tissue.Committed Progenitor Cells

[0166] In some embodiments, the regenerative cell population is a population of committed progenitor cells. Committed progenitor cells can be selected from a group comprising endothelial progenitor cells, hematopoietic progenitor cells, hematopoietic stem cells, neuronal progenitor cells, mesenchymal stem cells, mesenchymal stromal cells, or very small embryonic-like stem cells.Endothelial Progenitor Cells

[0167] Committed endothelial progenitor cells can be purified from bone marrow, peripheral blood, or the peripheral blood of a patient whose committed endothelial progenitor cellsVABIO.002WO2 PCT APPLICATIONhave been mobilized by the administration of a mobilizing agent or therapy. Nonlimiting examples of mobilizing agents include G-CSF, M-CSF, GM-CSF, 5-FU, IL-1, IL-3, kit-L, VEGF, Fit- 3 ligand, PDGF, EGF, FGF-1, FGF-2, TPO, IL- 11, IGF-1, MGDF, NGF, HMG CoA reductase inhibitors, and small molecule antagonists of SDF-1. In some embodiments, mobilization therapy can include exercise, hyperbaric oxygen therapy, autohemotherapy via ex vivo ozonation of peripheral blood, and induction of SDF-1 secretion in an anatomical area outside the bone marrow. In some embodiments, committed endothelial progenitor cells express an at least one marker selected from a group comprising: CD31, CD34, AC133, CD146, or flkl.Hematopoietic Progenitor Cells

[0168] In some embodiments, committed hematopoietic cells are purified from bone marrow or peripheral blood. In some embodiments, committed hematopoietic cells are purified from the peripheral blood of a patient whose commited hematopoietic progenitor cells have been mobilized by the administration of an at least one mobilizing agent or therapy. Nonlimiting examples of a mobilizing agent include G-CSF, M-CSF, GM-CSF, 5-FU, IL-1, IL-3, kit-L, VEGF, Fit- 3 ligand, PDGF, EGF, FGF-1, FGF-2, TPO, IL-11, IGF-1, MGDF, NGF, HMG CoA reductase inhibitors, and small molecule antagonists of SDF-1. In some embodiments, mobilization therapy can include exercise, hyperbaric oxygen therapy, autohemotherapy by ex vivo ozonation of peripheral blood, induction of SDF-1 secretion in an anatomical area outside the bone marrow, or any combination thereof. In some embodiments, committed hematopoietic progenitor cells express the marker CD 133 and / or CD34.Conditioned / Culture Media

[0169] In some embodiments, conditioned media is used in conjunction with or as a substitute for extracellular vesicles. In some embodiments, extracellular vesicles are administered together with a regenerative cell population. In some embodiments, extracellular vesicles can be isolated from regenerative cells or fluids of autologous, allogeneic, or xenogeneic origin.Exosomes

[0170] In some embodiments, exosomes can express at least one tetraspanin molecule. Nonlimiting examples of a tetraspanin molecule include TSPAN1, TSPAN2, TSPAN3, TSPAN4VABIO.002WO2 PCT APPLICATION(also known as NAG-2), TSPAN5, TSPAN6, TSPAN7 (also known as CD231, TALLA-1, Al 5), TSPAN8 (also known as CO-029), TSPAN9 (also known as NET-5), TSPAN10 (also known as OCULOSPANIN), TSPAN11 (also known as CD151-like), TSPAN12 (also known as NET-2), TSPAN13 (also known as NET-6), TSPAN14, TSPAN15 (also known as NET-7), TSPAN16 (also known as TM4-B), TSPAN17, TSPAN18, TSPAN19, TSPAN20 (also known as UPlb, UPK1B), TSPAN21 (also known as UPla, UPK1A), TSPAN22 (also known as RDS, PRPH2), TSPAN23 (also known as R0M1), TSPAN24 (also known as CD151), TSPAN25 (also known as CD53), TSPAN26 (also known as CD37), TSPAN27 (also known as CD82, KAI1), TSPAN28 (also known as CD81, TAPA-1), TSPAN29 (also known as CD9), TSPAN30 (also known as CD63), TSPAN31 (also known as SAS), TSPAN32 (also known as TSSC6), and TSPAN33. In some embodiments, exosomes express at least one tetraspanin molecule, optionally which include at least 1, 2, 3, 4, or 5 of: CD9, CD63, CD151, Tspan9, or Tspan32.

[0171] In some embodiments, exosomes are selected for expression of one or more markers associated with regenerative activity. In some embodiments, exosomes may express an at least one marker. Nonlimiting examples of markers include tetraspanins, CD9, CD63, CD81, ALIX, ESCRT-III components such as CHMP4B, ESCRT-I components such as TSG101, Rab GTPases, flotillin, heat shock proteins such as Hsp60, Hsp70, or Hsp90, growth factors such as PDGF-BB, FGF-2, VEGF-A, or HGF, cytokines such as TGF-P, microRNAs such as miR-1246, miR-21, miR-140-5p, miR-92a-3p, and miR-100-5p, pluripotency-related markers such as PODXL, specific surface markers based on stem cell source, such as BMSC-exos — which express markers such as CD63, CD9, and TSG101 — ADSC-exos — which express markers such as CD9, CD63, and CD81 — HFMSC-exos — which express markers such as CD13, CD9, CD63, CD105, CD81, CD29, CD44, CD49e, and SSEA-4 — and iPSC-exos — which express markers such as CD9, TSG101, and SSEA-1. In some embodiments, exosomes may express at least 1, 2, 3, 4, 5, 6 or 7 of the marker: CAV1, FLOTl, FLOT2, PROMI, CD63, CD9, and LAMP1. In some embodiments, exosomes are about 20 nm to about 300 nm in diameter, or any value that is between about 20 nm and about 300 nm in diameter. In some embodiments, exosomes have a diameter ranging from about 30 nm to about 150 nm.MicrovesiclesVABIO.002WO2 PCT APPLICATION

[0172] In some embodiments, microvesicles have a diameter ranging from about 100 nm, about 150 nm, about 200 nm, about 250 nm, about 300 nm, about 400 nm, about 500 nm, about 600 nm, about 700 nm, about 750 nm, about 800 nm, about 900 nm, about 1000 nm, or any value that is between about 100 nm and about 1000 nm. In some embodiments, microvesicles have an irregular shape. In some embodiments, microvesicles express tissue factor. In some embodiments, microvesicles express the alpha3betal integrin. In some embodiments, microvesicles express markers found on the cell membrane of their cell of origin. In some embodiments, microvesicles express E-selectin,Apoptotic Bodies

[0173] In some embodiments, apoptotic bodies include phosphatidyl serine on their membranes. In some embodiments, apoptotic bodies have an irregular shape. In some embodiments, apoptotic bodies have a diameter ranging from about 1 to about 5 um, or any value that is between 1 um and about 5 um. In some embodiments, apoptotic bodies are about 50 nm to about 5000 nm, or any value that is between about 50 nm and about 5000 nm, in diameter. In some embodiments, apoptotic bodies are bound to histones.Stem Cells

[0174] In some embodiments, mesenchymal stromal cells are derived from Bone marrow (BM-MSCs), Adipose tissue (AD-MSCs), Umbilical cord (UC-MSCs), Dental pulp, Placenta, Synovial Fluid, or other biomaterials. In some embodiments, mesenchymal stromal cells surface markers are positive for at least one of: CD73, CD90, CD105, or any combination thereof. In some embodiments, mesenchymal stromal cells surface markers are negative for at least one of: CD34, CD45 (hematopoietic markers), CD14, CD19, HLA-DR, or any combination thereof.

[0175] In some embodiments, stem cells are derived from a donor. In some embodiments, induced pluripotent stem cells (iPSCs) are derived from a patient / subject. In some embodiments, stem cells, iPSCs, and tlieir-derived extracellular vesicles are naturally sourced. In some embodiments, stem cells are primed by an at least one exogenous hormone. Non-limiting examples of exogenous hormones include estrogen, progesterone, Follicle-Stimulating Hormone (FSH), Luteinizing Hormone (LH), Human Chorionic Gonadotropin (hCG), and androgens. In some embodiments, priming cells increases the regenerative properties of stem cells, iPSCs, andVABIO.002WO2 PCT APPLICATIONtheir derived extracellular vesicles, such as: by reducing apoptosis and increasing proliferation, differentiation, migration, angiogenesis, and immuno-regulation.

[0176] In some embodiments stem cells, including induced pluripotent stem cells (iPSCs), can be primed with exogenous inflammatory molecules (such as interferon gamma (IFN-y), interleukin (IL)-la / (3, IL-25, IL-6, TNF-a, and IL-17) to enhance the anti-inflammatory properties of these cells and their derived extracellular vesicles. In some embodiments stem cells, including induced pluripotent stem cells (iPSCs), can be primed in a hypoxic environment to enhance the angiogenic properties of these cells and their derived extracellular vesicles. In some embodiments stem cells, iPSCs, and their-derived extracellular vesicles can be cultured in 2D or 3D systems to increase yield and regenerative properties.EXAMPLES

[0177] The following examples are given for the purpose of illustrating various embodiments of the disclosure and are not meant to limit the present disclosure in any fashion. One skilled in the art will readily appreciate that the present disclosure is well adapted to carry out the objects and obtain the ends and advantages mentioned and those objects, ends, and advantages inherent herein. Changes therein and other uses which are encompassed within the spirit of the disclosure, as defined by the scope of the claims, will occur to those skilled in the art. Examples of some embodiments containing specific biomaterials, manufacturing methods for these embodiments, and testing these embodiments for desired physical properties and device efficacy are illustrative and not intended to limit the present disclosure in any fashion.

[0178] Aspects of the present disclosure further relates to genitourinary administration of stem cells, iPSCs, and their derivative extracellular vesicles, or any combination thereof to a subject. In some embodiments, the subject has a disease, disorder, or condition related to the genitourinary system. In some embodiments, the disease, disorder, or condition comprises degeneration and / or damage to the genitourinary system. In some embodiment, the subject has at least one of: Genitourinary Syndrome of Menopause (GSM), Postpartum Vaginal Damage (PVT)), Mayer-Rokitansky-Kiister-Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), pelvic floor dysfunction, vaginitis, Ulis, and female infertility.

[0179] In some embodiments, stem cell-derived extracellular vesicles (EV s) are used either individually or in combination with stem cells. In some embodiments, the genitourinaryVABIO.002WO2 PCT APPLICATIONadministration of stem cells or derivatives thereof occurs in the vagina. In some embodiments, the treatment is administered through a biocompatible vaginal suppository.

[0180] In some embodiments, the method of preparation includes loading at least one of an iPSC, stem cell, extracellular vesicle, or any combination thereof, into a biocompatible suppository.

[0181] Also disclosed herein is a method of treating a subject in need thereof, the method comprising administering the biocompatible suppository to the subject. In some embodiments, the suppository is administered into the vaginal cavity. In some embodiments, the biocompatible suppository degrades when in contact with the subject. In some embodiments, during the gradual degradation process of the suppository, the at least one iPSC, stem cell, extracellular vesicle, or their combination are released into the inner wall of the female genitourinary tract. In some embodiments, iPSCs, stem cells, and extracellular vesicles gradually release chemokines, cytokines, hormones, growth factors, proteins, microRNA, mRNAs, bioactive lipids, and / or other biomolecules to the local environment. In some embodiments, this treatment reduces cell apoptosis and / or increases at least one of the following: cell proliferation, differentiation, migration, angiogenesis, immune regulation, rebuilding cell-to-cell contact, rejuvenating cells, repairing damage, or regenerating the vaginal and urethral inner walls, or any combination thereof. In some embodiments, these effects can restore the function of the lower genital-urinary tract and affected areas of the pelvic floor, alleviating genitourinary conditions, degeneration, and / or damage.Example 1: Preparation of IPSCs

[0182] In this example, vaginal suppositories were made from chitosan HC1, crosslinking agent’s collagen type I, carboxymethyl cellulose, and mucolytic enzyme DNase I

[0183] Induced pluripotent stem cells (IPSCs) are made from patient’s cell lines by culturing in Iscove’s modified Dulbecco’s medium (1MDM) supplemented with 10% (v / v) fetal bovine serum, penicillin (100 U / ml), and streptomycin (100 mg / ml; culture conditions are at 5% in CO2, the temperature was set to 37°C, cultured for 2 days and passaged once, the stem cells passaged 3-5 generations are subjected to subsequent processing. The culture medium (CM) is then collected in serum-free culture medium from stem cells, and the extracellular vesicles (EVs)VABIO.002WO2 PCT APPLICATIONare collected in serum-free culture medium by differential ultracentrifugation from the culture medium.

[0184] A suppository is made from mixture of chitosan HC1 and cross-linking agent’s collagen type I and carboxymethyl cellulose to prepare scaffolds with weight ratios of chitosan to cross-linking agent as follows 40:30:30, and 1-lOOug / ml DNase I through lyophilization. The porous and sponge chitosan gel carrier can hold at least one of: CM, EVs, stem cells, or any combination thereof. In some embodiments, the CM, EVs, stem cells, or any combination thereof, is loaded to the suppository and placed around or under the inner wall of the genitourinary tract. During the gradual degradation process of the suppository, stem cells continue proliferation and at same time, CM / EVs gradually release chemokines, hormones, growth factors, proteins, microRNA, mRNAs and bioactive lipids to the local tissue. These effects can rebuild cell-cell contact and rejuvenate the cells, repair any damage, and promote regeneration of the vaginal and urethral inner wall.

[0185] In some embodiments of the present disclosure, stem cells are derived from a donor using the standard protocol. In some embodiments, iPSCs are derived from a patient. In some embodiments, at least one of: stem cells, iPSCs, extracellular vesicles, or any combination thereof are locally sourced. In some embodiments, at least one of: stem cells, iPSCs, are primed by an at least one hormone. Non-limiting examples of priming hormones include estrogen, progesterone, relaxin, Follicle-Stimulating Hormone (FSH), Luteinizing Hormone (LH), Human Chorionic Gonadotropin (hCG), and androgens. In some embodiments, at least one of: stem cells, iPSCs, are primed by an at least one inflammatory molecule. Non-limiting examples of inflammatory molecules include interferon gamma (IFN-y), interleukin (IL)-la / p, IL-25, IL-6, TNF-a, IL23 and IL- 17. In some embodiments, at least one of: stem cells, iPSCs, are primed with hypoxia. In some embodiments, at least one of: stem cells, iPSCs are cultured in a 2D and / or 3D system to increase yield. In some embodiments, at least one of: stem cells, iPSCs, extracellular vesicles, or any combination thereof, are loaded onto a proprietary vaginal drug delivery suppository to be administered to a subject.Preparation of HUCMSCs-CM

[0186] HUCMSCs were cultured in 10 cm plates in DMEM / F12 supplemented with 10% fetal bovine serum (FBS), GlutaMAX, penicillin-G and streptomycin, and basic fibroblastVABIO.002WO2 PCT APPLICATIONgrowth factor (all from Gibco). Upon reaching 80% confluence, cells were detached by TrypLETM Express (Invitrogen) and passaged at the ratio of 1:3. For the collection of HUCMSC-CM, HUCMSC at passages 4-6 were used. At ~80% confluence, cells were washed three times with phosphate-buffered saline (PBS) and cultured with 6 ml of serum-free DMEM / F12 for 24 h. Afterwards, the culture medium was collected. The supernatant was centrifuged at 1500 rpm for 5 min to remove any cell material, and then the CM were pooled together and keep in 4 degrees for future use.Preparation of HUMSCs-EVs

[0187] The primary generation of HUCMSCs was offered by HUCMSCs core in the US, and cultured in a-minimum essential medium (a-MEM, Gibco, Gaithersburg, MD, USA) with 10% fetal bovine serum (FBS) (Sijiqing, Hangzhou, Zhejiang, China) and 1% penicillin / streptomycin (Gibco) at 37C in 5% CO2 humidified air. Three to eight passages of HUCMSCs were cultured to produce EVs. When the density of HUCMSCs reached 70%-80% confluency, 10% FBS was replaced by 0.5% of bovine serum albumin (BSA) (Sigma- Aldrich, StLouis, MO, USA) for 24-48 h (Fig. 1A). When HUCMSCs confluence reached 90% or more, the serum-free culture medium was collected and stored at 4C, and then EVs were isolated within 24 h by differential ultracentrifugation. Briefly, the cell supernatants were obtained through centrifugation at 300 g for 10 min and 2,000 g for 20 min to remove cells and debris, cell-free supernatants were then centrifuged at 100,000 g (Beckman Coulter Optima L-100XP ultracentrifuge) for 70 mm at 4C, the sediments were resuspended in phosphate buffer saline (PBS) (Hyclone, Logan, UT, USA) and submitted to a second and third 100,000 g ultracentrifugation at the same conditions. EV-associated proteins were determined by the Bradford method to estimate the amount of EVs17, About 600 ml serum-free culture medium of human umbilical mesenchymal stromal cells (hucMSCs) were used for collecting EVs, and 162.7 ± 17.5 mg EVs proteins were isolated in every 106HUCMSCs (about 10 ml of MSCs’ serum-free culture medium).Preparation of scaffolds:

[0188] A mixture of chitosan HC1 and cross-linking agent’s collagen type I and carboxymethyl cellulose (all from Sigma Aldrich, USA) to prepare scaffolds with weight ratios of chitosan to cross-linking agent as follows 40:30:30, then add l-100ug / ml DNase Ito the mixture,VABIO.002WO2 PCT APPLICATIONthe solution containing the cross-linking agent was added portion wise to chitosan HC1 solution during stirring on magnetic stirrer at 200 rpm (IKA® C-MAG HS 10 digital, Germany) to avoid formation of sticky masses. The resulting turbid mixture was poured into molds (1 cm long and 0.3cm in diameter), and placed in a refrigerator at -80 °C for 48 h. Then the mixture was lyophilized using Christ freeze dryer (ALPHA 2-4 LD plus, Germany) for 24 h. All scaffolds were sterilized by gamma radiation at 10 kGy (1.88 kGy / h) to prevent any contamination.Loading scaffolds with cells, CM, and EVs:

[0189] hucMSCs were purchased from stem cell bank. Frozen hucMSCs between P3 and P9 were freshly seeded in 10-cm culture dishes (1×106cells per dish) in Iscove’s modified Dulbecco’s medium (IMDM, Genom, China) supplemented with 10% (v / 'v) fetal bovine serum (FBS, Gibco, USA), penicillin (100 U / ml), and streptomycin (100 mg / ml; Gibco, USA), Briefly, the phenotypes of hucMSCs were specifically identified by FACS, and the osteogenic and adipogenic capacities of the mesenchymal stromal cells were assessed with a MesenCult Osteogenic Stimulatory Kit and a MesenCult Adipogenic Differentiation Kit (STEMCELL Technologies Inc., Canada).

[0190] hucMSCs at the third passage were trypsinized, centrifuged and reconstituted in new media. The cells number was adjusted to 10,000 cells per 50 pL of the media. Resuspend 500mg EVs proteins with 50ml CM, make the concentration of CM and EVS to 10:1 ratio. The scaffolds with 1 *0.3 cm dimensions were then loaded with 1ml of the CM and EV suspension on the top of each of the sterilized scaffold first, then load with 100,000 hucMSCs by dropping 0.5ml of the cell’s suspension on the top of each of the sterilized scaffold, the loading of stem cells and CM, EVs was done in aseptic conditions in a laminar flow hood equipped with UV light, then encapsuled and keep in 4 degree before use.Example 2: Preparation of exosomes from human umbilical mesenchymal stromal cells (huMSC-EV s )

[0191] Human umbilical mesenchymal stromal cells (huMSCs) were cultured in serum- free culture medium; the seeding number started with 5 × 105cells / 10 cm dish. At approximately 50% confluence, the medium was replaced with fresh medium containing estrogen (100 nM) for 48 hours. The culture dishes were washed three times with PBS, and serum-freeVABIO.002WO2 PCT APPLICATIONmedium was added. The cells were cultured for another 48 hours. The supernatants were obtained through centrifugation at 300 units of gravity (g) for 10 minutes, followed by centrifugation at 2,000 g for 20 minutes to remove cells and debris. Cell-free supernatants were then centrifuged at 100,000 g (Beckman Coulter Optima L-100XP ultracentrifuge) for 70 minutes at 4°C. The sediments were resuspended in phosphate-buffered saline (PBS) (Hyclone, Logan, UT, USA) and subjected to a second and third 100,000 g ultracentrifugation under the same conditions. Extracellular vesicle-associated proteins were determined by the Bradford method to estimate the amount of EVs. EVs were stored in -80 °C. Approximately 162.7 ± 17.5 mg of EV proteins were isolated from every 106huMSCs,

[0192] Loading huMSC-EVs into vaginal suppository. First, lOOpg of hucMSC-EVs were resuspended with 1 mL of PBS. Then, the vaginal suppository (roughly cylindrical scaffold) was loaded with 1 mL of the huMSC-EVs suspension on top of each of the sterilized scaffolds. The loading of huMSC-EVs was performed in aseptic conditions within a laminar flow hood equipped with UV light (ThermoFisher Scientific, USA). The vaginal suppository was then encapsulated and stored at 4°C prior to use.Example 3: Preparation of small extracellular vesicles (sEVs) from young sheep blood

[0193] Young sheep blood was extracted by aseptic venipuncture and collected into a vacuum tube (~5 mL) containing ethylenediaminetetraacetic acid (EDTA) as an anticoagulant. Plasma was obtained by centrifuging whole blood at room temperature for 15 minutes at 1,600 g. The purified plasma samples were immediately centrifuged to obtain the purified sEVs, and these were then stored at -80 °C. Plasma sEVs were isolated by ultracentrifugation. In brief, the plasma was diluted with an equal volume of phosphate-buffered saline (PBS) and subjected to a series of centrifugation steps at different speeds and durations. Specifically, centrifugation was performed at 500 g for 5 minutes at 4°C, followed by 3,000 g for 25 minutes at 4°C, 12,000 g for 60 minutes at 4°C, and, finally, 120,000 g for 70 minutes at 4°C (polycarbonate tubes, 355630, Beckman Coulter; MLA-55 rotor, Optima MAX-XP, Beckman Coulter). The resulting sEV pellet was collected, and the remaining supernatant was further processed using the Total Exosome Isolation Kit (from plasma) (Invitrogen, 4484450) according to the manufacturer’s instructions. In brief, 0.2 volumes of Exosome Precipitation Reagent were added to the supernatant, which was then incubated at room temperature for 10 minutes. Subsequently, the sEVs and the supernatant wereVABIO.002WO2 PCT APPLICATIONseparated via centrifugation at 10,000 g for 30 minutes at 4°C, with the resulting sEVs pellets being collected and resuspended in PBS. The yield of sEVs was assessed by quantifying the total protein content using the Bicinchoninic Acid (BCA) Kit (Thermo Fisher Scientific). sEVs were stored in -80 °C. Approximately 1,000 pg of total protein content of sEVs could be obtained from 1 mL of plasma.

[0194] Loading sEVs into vaginal suppository; First, 100pg of sEVs were resuspended with 1 mL of PBS. Then, the vaginal suppository (roughly cylindrical scaffold) was loaded with 1 mL of the sEV suspension on top of each of the sterilized scaffolds. The loading of sEVs was performed in aseptic conditions within a laminar flow hood equipped with UV light (ThermoFisher Scientific, USA). The vaginal suppository was then encapsulated and stored at 4°C prior to use.Conclusions:

[0195] Each of Examples 1, 2 and 3 will be tested in the post-menopausal rat model and the expected results will include one or more of: decreased inflammation, increased angiogenesis in vagina, proliferation of vaginal epithelial cells, enhanced intercellular adhesion, thickening of the epithelial layer, increased vagina healthy score, and healing of the vaginal wound and erosive surface to reverse the atrophy, without changing hormone levels in rats.Example 4: Administering Therapeutic Compounds to Mammals

[0196] Forty adult, female, specific pathogen free (SPF) Wistar rats, weighing about 250- 300 g were fed with standard rat food and water. The rats were allowed to drink ad libitum and were maintained on a 12 hour light / 12 hour darkness cycle. Eight rats were randomly assigned to undergo sham debulking (Sham group) and the rest were established as a bilateral ovariectomy (OVX) animal model to simulate menopause. A 40 mg / kg intraperitoneal injection of 1% pentobarbital sodium was used to anesthetize the rats. The rats’ ovaries were removed, and the statuses of the rats were observed daily. The presence of discharge (blood) from the vaginal opening, and redness and congestion of the vaginal mucosa was recorded. After recovery from surgery, the vaginal pH of the rats was measured for five consecutive days and vaginal smears were performed by dipping a sterile cotton swab in saline, simultaneously rotating and inserting the swab slowly into the vagina of the rats, gently removing the swab with a few rotations, and applying the residue to a clean slide that had been dried in 95% ethanol. Routine hematoxylin eosinVABIO.002WO2 PCT APPLICATIONstaining (HE staining) was performed on the swab residue to observe the keratinization and shedding of epithelial cells. This testing began after the rats’ vaginas became congested, red, and swollen, and began discharging a thick secretion, indicating that both ovaries had been completely removed and that the rats were in hypoestrogenic states.

[0197] Each group of 8 successfully molded rats was randomly divided into four groups namely: model control group (M); blank suppository group (B); positive control (vaginal ET) group (P); and SCEV suppository administration group (SCEV). Vaginal administration, equal volume of 1 capsule / 0.2 g / each, was administered once, then an examination was performed after 4 weeks.Cell Observation Count of Vaginal Iavage Smears:

[0198] Before and 4 weeks after the administration of the drug, the vaginal health score was performed on a scale of 1 to 4 with higher scores associated with less atrophy. pH, discharge, epithelial mucosa, wetness, vaginal folds, and mucosal color were used as indicators. Vaginal lavage smears and HE staining were performed to observe the morphology of exfoliated cells, and high magnification microscopy was used to count exfoliated epithelium, inflammatory cells, and epithelial classification.Intravaginal Microbiological Testing:

[0199] Before and 4 weeks after the administration of the drug, the vaginal lavage was collected for culture and detection of lactobacilli; the amount of lactobacilli in the vaginal secretions was observed by staining with Gram stain.ELISA Kit to Determine Sex Hormone Levels

[0200] 24 h after the dose, blood was collected from the abdominal aorta, and the serum levels of Estradiol (E2) and Follicle-stimulating Hormone (FSH) were measured according to ELISA kits.Organs of the Uterus and Vagina Index

[0201] After 4 weeks of dosing, the animal was executed, the upper end of the urethra was taken at about 1 cm, and the skin and abdominal wall were cut longitudinally at about 3 cm. TheVABIO.002WO2 PCT APPLICATIONuterus and vagina were peeled out, and the blood was flushed out with saline. Excess water was blotted out with filter paper, and the animal was weighed together with an electronic balance to calculate the organ coefficient and perform pathological histological observation with the formula: Organ index = organ weight (mg) / body weight (g).Determination of Glycogen Content

[0202] After 4 weeks, the vagina was taken 0.5 cm fixed in 10% formalin, dehydrated and embedded, oxidized with oxidant, stained sequentially with Schiff and hematoxylin stains, and differentiated and returned to blue. PAS (Periodic Acid-Schiff’ s reaction) staining was carried out to observe the glycogen content in the vaginal tissue.Histomorphological Observation of the Vagina

[0203] Vaginal tissue specimens were fixed in 10% formalin for 2 weeks, dehydrated in an alcohol series, xylene transparent, wax dipped, embedded and cut into 0.005 mm thick slices for HE staining to observe vaginal morphology and thickness in rats.Immunohistochemical Detection of EGF, E-Cadherin Protein Expression

[0204] Rat vaginal tissue was dipped in wax, embedded, and sectioned. After deparaffinisation, the tissue was incubated with hydrogen peroxide to quench the endogenous peroxidase activity and heated to inactivate the endogenous enzymes and biotin. Tissues were incubated with antibodies against the epidermal growth factor (EGF) and epithelial cadherin (E-cadherin) for 60 min at 37 °C. Reaction enhancers were added dropwise, secondary antibodies were incubated for 20 min, developed with 3,3-diaminobenzidine (DAB) for 5 min and finally counterstained with haematoxylin. The tissue was observed with a microscope and photographed. To assess protein expression, the intensity of DAB was analysed and scored using Image J. Three randomly selected fields of view were photographed under a light microscope for scoring and cells showing brown signals were defined as immunohistochemically positive. The intensity of staining was categorised as: negative 0, low positive 1, moderate positive 2 and high positive 3. The percentage of stained areas was 0 for 0 to 5%, 1 for 6% to 25%, 2 for 26% to 49% and 3 for >50%. The final score was measured by multiplying the combined scores of the two. A total score of 0 isVABIO.002WO2 PCT APPLICATIONconsidered negative, 1 to 3 as low expression, 4 to 6 as moderate expression and 6 to 9 as high expression.Real-Time PCR Assay to Detect the mRNA Expression of EGF and E-Cadherin

[0205] Total RNA from rat vaginal tissues of each group was collected and extracted with TRIzol reagent (Thermo Fisher Scientific, Shanghai, China). The concentration of RNA was detected by Multifunctional Enzyme Labeler and then total RNA was reverse transcribed into cDNA according to the steps of reverse transcription kit. Amplification conditions were: 95C for 10 min, 95 C for 10 s, 60 C for 30 s, 72 C for 30 s, 40 cycles; lysis curve analysis: 95 C for 10 s, step sampling, step temperature 0.5 C. The relative mRNA expression of EGF and E-cadherin in vaginal tissues of rats in each group was calculated using RN-ACTB as an internal reference.Results

[0206] There was no significant difference in body weight between the groups before surgery. After bilateral oophorectomy and before treatment administration, the sham-operated group (Sham) had normal fur color and body shape compared to the wild type group, whereas other groups showed coarse and lustrous coat color, redness and swelling of the vaginal opening, increased discharge, frequent itchy movements, and significantly larger body shape, with a significant increase in body weight and higher pH.

[0207] After 4 weeks of treatment administration, both the positive control group (P) and the stem cells and extracellular vesicles suppository group (SCEV) showed reduced redness and swelling, decreased discharge, reduced inflammatory conditions, significantly lower pH than in the Sham group, and significantly higher vaginal health scores. However, the SCEV group had a more pronounced effect than the P group.

[0208] HE staining showed that the squamous epithelial layer in the vaginal tissue of Sham group rats was structurally intact, with a single layer of columnar epithelium covering the endothelial surface and tightly connected. Reduction in the number of squamous epithelial layers and thinning of the vaginal wall were observed in the B group. In contrast, the number of vaginal squamous epithelium, the number of mucosal epithelial layers, and the thickness of the vaginal wall were increased in the dosed P and SCEV rats compared to the B rats, with the most pronounced increase in the SCEV group.VABIO.002WO2 PCT APPLICATION

[0209] The rats in the Sham group had an estrous phase with a large number of epithelial cells (including unkeratinized cells, incompletely keratinized cells, and keratinized exfoliated cells) in the vagina, and almost no inflammatory cell infiltration. In the other groups, the ovaries were successfully removed for modeling, and inflammatory cell infiltration was evident in the mucosal layer and submucosal layer or the whole layer of the vagina, and epithelial cells were significantly reduced. The number of inflammatory cells in the vagina of the rats in the P and SCEV groups was significantly reduced, and the number of epithelial cells (including unkeratinized cells, incomplete keratinized cells, and keratinized exfoliated cells) was significantly higher than that in the B group. Once again, the effect of the SCEV group was better than that of the P group. The vaginal index of the rats in the B groups was significantly lower than that of the Sham group, which indicated that the effect of ovarian removal was significant. The vaginal index of the two treatment-administered groups each increased by different degrees after treatment, and the atrophy was improved.

[0210] After ovarian removal, serum levels of E2 were significantly lower and FSH levels were significantly higher in the B group compared to the Sham group, which indicated a trend of decreased estradiol (E2) and increased follicle-stimulating hormone (FSH) during menopause after successful modeling. Serum E2 levels were significantly higher in group P than in group B after administration and the serum FSH level of group P was significantly lower than that of group B after administration. This indicated that the vaginal estrogen therapy in the P group achieved its therapeutic effect on atrophic vaginitis by regulating hormone levels. In contrast, there was no significant change in the SCEV groups, which indicated that the stem cells and extracellular vesicles administration group did not contain hormones and did not affect the hormone levels of the rats themselves.

[0211] Normally, estrogen stimulates the epithelial cells to produce glycogen. This glycogen is converted to lactic acid by lactic acid bacteria such as lactobacilli, in the vagina, resulting in an acidic pH. Compared to the Sham group, glycogen levels were significantly reduced in the B group and glycogen levels were increased in both the P and SCEV groups after dosing. Gram staining indicated the number of lactobacilli found in vaginal secretions. A significant decrease in vaginal lactobacilli in the B group after surgery was observed compared to the Sham group. Additionally, Gram staining showed a significant increase in lactobacilli in the SCEV group after administration of the treatment.VABIO.002WO2 PCT APPLICATION

[0212] The immunohistochemical results of rat vaginal tissue sections were scored according to the immunohistochemical scoring criteria. EGF protein expression in the Sham group, P group, and SCEV group were significantly different compared with the model B group. The SCEV group had higher EGF expression than the P group. Epithelial calcium adhesin (E-cadherin) protein expression in Sham group and SCEV group were significantly different compared with the model B group. An increase in the intensity of E-cadherin on the plasma membrane of epithelial cells was observed in SCEV group. This indicates an increased expression of cell adhesion proteins between adjacent cells. Additionally, the mRNA expression of EGF and E-cadherin was significantly higher in the Sham, P and SCEV groups compared to the B group.Conclusions

[0213] Stem cells, and stem cell-derived extracellular vesicles, were shown to decrease inflammation, increase the expression of EGF and E-cadherin in vaginal epithelial cells, promote the proliferation of vaginal epithelial cells, enhance intercellular adhesion, thicken the epithelial layer, recover the vagina’s self-cleaning ability, and heal the vaginal wound and erosive surface to improve atrophy. Furthermore, the composition as part of a vaginal suppository demonstrated a high safety profile without changing hormone levels in rats.Example 5: Non-limiting Example Uses and Methods of Treating

[0214] The composition of the present disclosure has many potential uses in treating diseases, disorders, or conditions associated with the female genitourinary tract. In some embodiments, the disease, disorder, or condition comprises Postpartum Vaginal Damage (PVD), Mayer-Rokitansky-Kuster-Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), female infertility caused by vaginal inflammation, immuno-unbalance of the uterus, vaginal dryness, vaginal damage, vaginal inflammation, vaginal wall thinning, or any combination thereof.

[0215] In some embodiments of the invention, the composition of any one of the present disclosure is administered to a subject for therapeutic use. In some embodiments, the composition is directly injected into the subject. In some embodiments, the composition is injected together with cellular supernatants and / or with extracellular vesicles. In some embodiments the composition is delivered using a suppository or suppository-like device. In some embodiments,VABIO.002WO2 PCT APPLICATIONthe composition comprises an at least one stem cell and / or an at least one stem cell-derived extracellular vesicles.

[0216] In some embodiments, the composition is loaded into a biocompatible suppository, wherein the suppository is designed to degrade / dissolve when inserted into the subject. In some embodiments, the composition is released into surrounding tissue once the suppository is dissolved.

[0217] In some embodiments, the composition comprises a therapeutic regenerative cell and / or a therapeutic extracellular vesicle. In some embodiments, the extracellular vesicle is derived from the regenerative cell. In some embodiments, the vesicle is isolated from the extracellular or intracellular supernatant of the regenerative cell. In some embodiments, the vesicle is further chemically modified.Certain Inventive Aspects

[0218] The following are numbered aspects of the disclosure:

[0219] 1. A first aspect comprising a method of preventing, inhibiting, or treating Genitourinary Syndrome of Menopause (GSM), Postpartum Vaginal Damage (PVD), Mayer- Rokitansky-Kuster -Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), and female infertility caused by vaginal inflammation, or immuno-unbalance of the uterus, comprising the steps of: a) identifying a patient at risk, or suffering from these diseases; b) isolating stem cells or iPSCs from a donor or a patient; c) priming stem cells or iPSCs with hormones, or hypoxia, or pro-inflammatory molecules to increase efficacy; d) isolating extracellular vesicles from primed stem cells and iPSCs; e) loading cells and / or extracellular vesicles onto a biocompatible suppository; and f) applying the suppository into the vaginal tract in a manner to allow for the gradual degradation and release of process of materials, such as cytokines, chemokines, growth factors, proteins, microRNA, miRNAs, mRNAs and bioactive lipids to the local environment allowing for re-establishment of cell-cell contact, immune balance, and rejuvenation of cells including the repair of damage and promotion of regeneration of vaginal and urethral inner wall, therefore restore the function of lower genital-urinary tract, alleviate these diseases.

[0220] 2. A second aspect comprising a method of preventing, inhibiting or treating Genitourinary Syndrome of Menopause (GSM), Postpartum Vaginal Damage (PVD), Mayer-VABIO.002WO2 PCT APPLICATIONRokitansky-Küster-Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), and female infertility caused by vaginal inflammation, or immuno-unbalance of the uterus, comprising the steps of: a) identifying a patient at risk of, or suffering from these diseases; b) isolating small extracellular vesicles from regenerative cells or fluids of autologous, allogeneic, or xenogeneic origin; c) loading small extracellular vesicles into a biocompatible suppository; and d) applying the suppository into the affected areas of the vaginal tract and pelvic floor in a manner to allow for the gradual degradation and release of process of materials, sEVs, proteins, microRNA, mRNAs and bioactive lipids to the local environment allowing for re-establishment of cell-cell contact, immune balance and rejuvenation of cells including the repair of damage and promotion of regeneration of vaginal and urethral inner wall, therefore restoring the function of the pelvic floor, alleviating these diseases.

[0221] 3. The method of aspect 1 or aspect 2, wherein said extracellular vesicles are selected from a group comprising: a) exosomes; b) apoptotic bodies; c) microvesicles; d) free mitochondria; and e) vesicular structures.

[0222] 4. The method of aspect 3, wherein said exosomes express at least one tetraspanin molecule.

[0223] 5. The method of aspect 4, wherein said tetraspanin molecules are selected from a group comprising: a) CD9; b) CD63; c) CD151; d) Tspan9; and e) Tspan32.

[0224] 6. The method of aspect 3, wherein said exosomes are selected for expression of one or more markers associated with regenerative activity.

[0225] 7. The method of aspect 6, wherein said regenerative activity is the stimulation of angiogenesis.

[0226] 8. The method of aspect 6, wherein said regenerative activity is the reduction of fibrosis.

[0227] 9. The method of aspect 6, wherein said regenerative activity is the reduction of inflammation.

[0228] 10. The method of aspect 6, wherein said markers associated with regenerative activity are selected from a group comprising: a) a heat shock protein; b) a stress-associated protein; and c) a growth factor receptor.

[0229] 11. The method of aspect 10, wherein said heat shock protein is selected from a group comprising: a) calreticulin; b) hsp20; and c) hsp90.VABIO.002WO2 PCT APPLICATION

[0230] 12. The method of aspect 10, wherein the stress-induced protein is phosphatidylserine.

[0231] 13. The method of aspect 10, wherein said growth factor receptor is selected from a group comprising: a) IL-3 receptor; b) VEGF receptor; c) CXCL12; and d) c-met.

[0232] 14. The method of aspect 2, wherein said regenerative cells are stem cells.

[0233] 15. The method of aspect 14, wherein said stem cells are hematopoietic stem cells.

[0234] 16. The method of aspect 15, wherein said hematopoietic stem cells express markers selected from a group comprising: a) CD133; b) CD34; c) c-kit; d) IL-3 receptor; e) c-mpl; 0 c-met; g) stem cell factor receptor; and h) LAP.

[0235] 17. The method of aspect 16, wherein said hematopoietic stem cells are capable of regenerating hematopoiesis in an immunodeficient mouse.

[0236] 18. The method of aspect 14, wherein said stem cells are mesenchymal stem cells and / or mesenchymal stromal cells.

[0237] 19. The method of aspect 18, wherein said mesenchymal stem and / or stromal cell is preconditioned, stimulates enhanced production of therapeutic effects.

[0238] 20. The method of aspect 19, wherein the preconditioning is accomplished by treatment with conditions that induce cellular stress.

[0239] 21. The method of aspect 20, wherein said cellular stress is characterized by nuclear translocation of NF-kappa B by more than 10% compared to baseline.

[0240] 22. The method of aspect 20, wherein said cellular stress is characterized by nuclear translocation of NF-kappa B by more than 20% compared to baseline.

[0241] 23. The method of aspect 20, wherein said cellular stress is characterized by nuclear translocation of NF-kappa B by more than 50% compared to baseline.

[0242] 24. The method of aspect 20, wherein the cellular stress is characterized by nuclear translocation of HIF-la by more than 10% compared to baseline.

[0243] 25. The method of aspect 20, wherein the cellular stress is characterized by nuclear translocation of HIF-la by more than 20% compared to baseline.

[0244] 26. The method of aspect 20, wherein the cellular stress is characterized by nuclear translocation of HIF-la by more than 50% compared to baseline.VABIO.002WO2 PCT APPLICATION

[0245] 27. The method of aspect 20, wherein said cellular stress is characterized by nuclear translocation of heme oxygenase- 1 by more than 10% compared to baseline.

[0246] 28. The method of aspect 20, wherein said cellular stress is characterized by nuclear translocation of heme oxygenase- 1 by more than 20% compared to baseline.

[0247] 29. The method of aspect 20, wherein said cellular stress is characterized by nuclear translocation of heme oxygenase- 1 by more than 50% compared to baseline.

[0248] 30. The method of aspect 20, wherein said cellular stress is characterized by nuclear translocation of TyrRS by more than 10% compared to baseline.

[0249] 31. The method of aspect 20, wherein said cellular stress is characterized by nuclear translocation of TyrRS by more than 20% compared to baseline.

[0250] 32. The method of aspect 20, wherein said cellular stress is characterized by nuclear translocation of TyrRS by more than 50% compared to baseline.

[0251] 33. The method of aspect 20, wherein said stress is oxidative stress.

[0252] 34. The method of aspect 20, wherein said stress is hypotonic stress.

[0253] 35. The method of aspect 20, wherein said stress is hypertonic stress.

[0254] 36. The method of aspect 20, wherein said stress is radiation-induced stress.

[0255] 37. The method of aspect 20, wherein said stress is ultrasound-induced stress.

[0256] 38. The method of aspect 20, wherein said stress is inflammation-induced stress.

[0257] 39. The method of aspect 38, wherein said inflammation-induced stress is the result of activation of one or more toll-like receptors.

[0258] 40. The method of aspect 39, wherein said toll-like receptor is TLR4.

[0259] 41. The method of aspect 40, wherein said TLR4 receptor activator is betaglucan.

[0260] 42. The method of aspect 40, wherein the TLR4 receptor activator is low-molecular-weight hyaluronic acid.

[0261] 43. The method of aspect 40, wherein said TLR4 receptor activator is histone- DNA complexes.

[0262] 44. The method of aspect 43, wherein said histone-DNA complexes are derived from neutrophil extracellular traps.

[0263] 45. The method of aspect 39, wherein said TLR is TLR3.VABIO.002WO2 PCT APPLICATION

[0264] 46. The method of aspect 45, wherein said TLR3 receptor activator is Poly IC.

[0265] 47. The method of aspect 1 or claim 2, wherein said regenerative cell is selected from either alone or in combination with a group comprising: stem cells, committed progenitor cells, and differentiated cells.

[0266] 48. The method of aspect 47, wherein said stem cells are selected from a group comprising of: embryonic stem cells, cord blood stem cells, placental stem cells, bone marrow stem cells, amniotic fluid stem cells, neuronal stem cells, circulating peripheral blood stem cells, mesenchymal stem cells, mesenchymal stromal cells, germinal stem cells, adipose tissue derived stem cells, exfoliated teeth derived stem cells, hair follicle stem cells, dermal stem cells, parthenogenically derived stem cells, reprogrammed stem cells and side population stem cells.

[0267] 49. The method of aspect 47, wherein said embryonic stem cells are totipotent

[0268] 50. The method of aspect 49, wherein said embryonic stem cells express one or more antigens selected from a group consisting of: stage-specific embryonic antigens (SSEA) 3, SSEA 4, Tra-1-60 and Tra-1-81, Oct-3 / 4, Cripto, gastrin-releasing peptide (GRP) receptor, podocalyxin-like protein (PODXL), Rex-1, GCTM-2, Nanog, and human telomerase reverse transcriptase (hTERT).

[0269] 51. The method of aspect 48, wherein said cord blood stem cells are multipotent and capable of differentiating into endothelial, smooth muscle, and neuronal cells.

[0270] 52. The method of aspect 51, wherein said cord blood stem cells are identified based on expression of one or more antigens selected from a group comprising: SSEA-3, SSEA-4, CD9, CD34, c-kit, OCT-4, Nanog, and CXCR-4

[0271] 53. The method of aspect 52, wherein the cord blood stem cells do not express one or more markers selected from the group comprising CD3, CD34, CD45, and CD1 lb.

[0272] 54. The method of aspect 48, wherein said placental stem cells are isolated from the placental structure.

[0273] 55. The method of aspect 54, wherein said placental stem cells are identified based on expression of one or more antigens selected from a group comprising: Oct-4, Rex-1, CD9, CD13, CD29, CD44, CD166, CD90, CD105, SH-3, SH-4, TRA-1-60, TRA-1-81, SSEA-4, and Sox-2.

[0274] 56. The method of aspect 48, wherein said bone marrow stem cells comprise bone marrow mononuclear cells.VABIO.002WO2 PCT APPLICATION

[0275] 57. The method of aspect 52, wherein said bone marrow stem cells are selected based on the ability to differentiate into one or more of the following cell types: endothelial cells, smooth muscle cells, and neuronal cells.

[0276] 58. The method of aspect 57, wherein said bone marrow stem cells are selected based on expression of one or more of the following antigens: CD34, c-kit, flk-1, Stro-1, CD105, CD73, CD31, CD146, vascular endothelial-cadherin, CD133, and CXCR-4.

[0277] 59. The method of aspect 58, wherein said bone marrow stem cells are enriched for expression of CD 133.

[0278] 60. The method of aspect 48, wherein the amniotic fluid stem cells are isolated by introducing a fluid extraction means into the amniotic cavity under ultrasound guidance,

[0279] 61. The method of aspect 60, wherein said amniotic fluid stem cells are selected based on expression of one or more of the following antigens: SSEA3, SSEA4, Tra-1-60, Tra-1-81, Tra-2-54, HLA class I, CD13, CD44, CD49b, CD105, Oct-4, Rex-1, DAZL, and Runx-1.

[0280] 62. The method of aspect 60, wherein said amniotic fluid stem cells are selected based on the lack of expression of one or more of the following antigens: CD34, CD45, and HLA Class II.

[0281] 63. The method of aspect 48, wherein said neuronal stem cells are selected based on expression of one or more of the following antigens: RC-2, 3CB2, BLB, Sox-2, GLAST, Pax 6, nestin, Muashi-1, NCAM, A2B5, and prominin.

[0282] 64. The method of aspect 48, wherein said circulating peripheral blood stem cells are characterized by the ability to proliferate in vitro for a period of over 3 months.

[0283] 65. The method of aspect 48, wherein said circulating peripheral blood stem cells are characterized by expression of CD34, CXCR4, CD117, CD113, and c-met.

[0284] 66. The method of aspect 65, wherein said circulating peripheral blood stem cells lack substantial expression of differentiation-associated markers.

[0285] 67. The method of aspect 66, wherein said differentiation-associated markers are selected from a group comprising CD2, CD3, CD4, CD11, CDlla, Mac-1, CD14, CD16, CD19, CD24, CD33, CD36, CD38, CD45, CD56, CD64, CD68, CD86, CD66b, andHLA-DR.

[0286] 68. The method of aspect 48, wherein said mesenchymal stromal cells express one or more of the following markers: STRO-1, CD105, CD54, CD106, HLA-I markers, vimentin, ASMA, collagen- 1, fibronectin, LFA-3, ICAM-1, PECAM-1, P-selectin, L-selectin,VABIO.002WO2 PCT APPLICATIONCD49b / CD29, CD49c / CD29, CD49d / CD29, CD61, CD 18, CD29, thrombomodulin, telomerase, CD10, CD13, STRO-2, VCAM-1, CD146, and THY-1.

[0287] 69. The method of aspect 68, wherein said mesenchymal stromal cells do not express substantial levels ofHLA-DR, CD117, and CD45.

[0288] 70. The method of aspect 48, wherein said mesenchymal stromal cells are derived from a group selected from: bone marrow, adipose tissue, umbilical cord blood, placental tissue, peripheral blood mononuclear cells, differentiated embryonic stem cells, and differentiated progenitor cells,

[0289] 71. The method of aspect 48, wherein said germinal stem cells express markers selected from a group comprising: Oct4, Nanog, Dppa5, Rbm, cyclin A2, Tex18, Stra8, Dazl, beta1- and alpha6-integrins, Vasa, Fragilis, Nobox, c-Kit, Sca-1, and Rexl.

[0290] 72. The method of aspect 48, wherein said adipose tissue-derived stem cells express markers selected from a group comprising CD 13, CD29, CD44, CD63, CD73, CD90, CD 166, Aldehyde dehydrogenase (ALDH), and ABCG2,

[0291] 73. The method of aspect 71, wherein said adipose tissue-derived stem cells are a population of purified mononuclear cells extracted from adipose tissue capable of proliferating in culture for more than 1 month.

[0292] 74. The method of aspect 48, wherein said exfoliated teeth-derived stem cells express markers selected from a group comprising: STRO-1, CD146 (MUC18), alkaline phosphatase, MEPE, and bFGF.

[0293] 75. The method of aspect 48, wherein said hair follicle stem cells express markers selected from a group comprising: cytokeratin 15, Nanog, and Oct-4.

[0294] 76. The method of aspect 48, wherein said hair follicle stem cells are capable of proliferating in culture for a period of at least one month.

[0295] 77. The method of aspect 48, wherein said hair follicle stem cells secrete one or more of the following proteins when grown in culture: basic fibroblast growth factor (bFGF), endothelin-1 (ET-1), and stem cell factor (SCF).

[0296] 78. The method of aspect 48, wherein the dermal stem cells express markers selected from the group comprising CD44, CD13, CD29, CD90, and CD 105.

[0297] 79. The method of aspect 48, wherein said dermal stem cells are capable of proliferating in culture for a period of at least one month.VABIO.002WO2 PCT APPLICATION

[0298] 80. The method of aspect 48, wherein parthenogenically derived stem cells are generated by the addition of a calcium flux-inducing agent to activate an oocyte, followed by enrichment of cells expressing markers selected from a group comprising SSEA-4, TRA-1-60, and TRA-1-81.

[0299] 81. The method of aspect 48, wherein said reprogrammed stem cells are selected from a group comprising of: cells subsequent to a nuclear transfer, cells subsequent to a cytoplasmic transfer, cells treated with a DNA methyltransferase inhibitor, cells treated with a histone deacetylase inhibitor, cells treated with a GSK-3 inhibitor, cells induced to dedifferentiate by alteration of extracellular conditions, and cells treated with various combination of the mentioned treatment conditions.

[0300] 82. The method of aspect 48, wherein said nuclear transfer comprises introducing nuclear material to a substantially enucleated cell, said nuclear material deriving from a host whose genetic profile is sought to be dedifferentiated.

[0301] 83. The method of aspect 81, wherein said cytoplasmic transfer comprises introducing cytoplasm of a cell with a dedifferentiated phenotype into a cell with a differentiated phenotype, such that said cell with a differentiated phenotype substantially reverts to a dedifferentiated phenotype.

[0302] 84. The method of aspect 81, wherein the DNA demethylating agent is selected from the group comprising 5 -azacytidine, psammaplin, and zebularine.

[0303] 85. The method of aspect 81, wherein the histone deacetylase inhibitor is selected from a group comprising valproic acid, trichostatin A, trapoxin A, and depsipeptide.

[0304] 86. The side population cells of aspect 48, wherein said cells are identified based on the expression of the multidrug resistance transport protein (ABCG2) or their ability to efflux intracellular dyes, such as rhodamine- 123 and / or Hoechst 33342.

[0305] 87. The side population cells of aspect 86, wherein said cells are derived from tissues such as pancreatic tissue, liver tissue, smooth muscle tissue, striated muscle tissue, cardiac muscle tissue, bone tissue, bone marrow tissue, bone spongy tissue, cartilage tissue, liver tissue, pancreas tissue, pancreatic ductal tissue, spleen tissue, thymus tissue, Peyer's patch tissue, lymph nodes tissue, thyroid tissue, epidermis tissue, dermis tissue, subcutaneous tissue, heart tissue, lung tissue, vascular tissue, endothelial tissue, blood cells, bladder tissue, kidney tissue, digestive tract tissue, esophagus tissue, stomach tissue, small intestine tissue, large intestine tissue, adipose tissue,VABIO.002WO2 PCT APPLICATIONuterus tissue, eye tissue, lung tissue, testicular tissue, ovarian tissue, prostate tissue, connective tissue, endocrine tissue, and mesentery tissue.

[0306] 88. The method of aspect 48, wherein said committed progenitor cells are selected from a group comprising: endothelial progenitor cells, neuronal progenitor cells, and hematopoietic progenitor cells.

[0307] 89. The method of aspect 88, wherein said committed endothelial progenitor cells are purified from the bone marrow.

[0308] 90. The method of aspect 89, wherein said committed endothelial progenitor cells are purified from peripheral blood.

[0309] 91. The method of aspect 90, wherein said committed endothelial progenitor cells are purified from the peripheral blood of a patient whose committed endothelial progenitor cells are mobilized by administration of a mobilizing agent or therapy,

[0310] 92. The method of aspect 91, wherein said mobilizing agent is selected from a group comprising: G-CSF, M-CSF, GM-CSF, 5-FU, IL- 1, IL-3, kit-L, VEGF, Flt-3 ligand, PDGF, EGF, FGF-1, FGF-2, TPO, IL-11, IGF-1, MGDF, NGF, HMG CoA reductase inhibitors, and small molecule antagonists of SDF-I.

[0311] 93. The method of aspect 92, wherein said mobilization therapy is selected from a group comprising exercise, hyperbaric oxygen, autohemotherapy by ex vivo ozonation of peripheral blood, and induction of SDF-1 secretion in an anatomical area outside of the bone marrow.

[0312] 94. The method of aspect 88, wherein said committed endothelial progenitor cells express markers selected from a group comprising: CD31, CD34, AC133, CD146, and flkl.

[0313] 95. The method of aspect 88, wherein said committed hematopoietic cells are purified from the bone marrow.

[0314] 96. The method of aspect 91, wherein said committed hematopoietic progenitor cells are purified from peripheral blood.

[0315] 97. The method of aspect 96, wherein said committed hematopoietic progenitor cells are purified from the peripheral blood of a patient whose committed hematopoietic progenitor cells have been mobilized by the administration of a mobilizing agent or therapy.

[0316] 98. The method of aspect 97, wherein said mobilizing agent is selected from a group comprising: G-CSF, M-CSF, GM-CSF, 5-FU, IL-1, IL-3, kit-L, VEGF, Flt-3 ligand, PDGF,VABIO.002WO2 PCT APPLICATIONEGF, FGF-1, FGF-2, TPO, IL-11, IGF-1, MGDF, NGF, HMGCoA reductase inhibitors, and small molecule antagonists of SDF-1.

[0317] 99. The method of aspect 97, wherein said mobilization therapy is selected from a group comprising: exercise, hyperbaric oxygen, aut chemotherapy by ex vivo ozonation of peripheral blood, and induction of SDF-1 secretion in an anatomical area outside of the bone marrow.

[0318] 100. The method of aspect 95, wherein said committed hematopoietic progenitor cells express the marker CD133.

[0319] 101. The method of aspect 95, wherein said committed hematopoietic progenitor cells express the marker CD34.

[0320] 102. The method of aspect 1 or 2, wherein an antioxidant is administered at a therapeutically sufficient concentration to a patient in need thereof.

[0321] 103. The method of aspect 102, wherein said antioxidant is selected from a group comprising: ascorbic acid and derivatives thereof, alpha tocopherol and derivatives thereof, rutin, quercetin, hesperedin, lycopene, resveratrol, tetrahydrocurcumin, rosmarinic acid, Ellagic acid, chlorogenic acid, oleuropein, alpha-lipoic acid, glutathione, polyphenols, pycnogenol.

[0322] 104. The method of aspect 1 or aspect 2, wherein said extracellular vesicles are exosomes.

[0323] 105. The method of aspect 104, wherein said exosomes express markers selected from a group comprising: CAV1, FLOT1, FLOT2, and PR0M1.

[0324] 106. The method of aspect 104, wherein said exosomes express markers selected from a group comprising: CD63, CD9, and LAMP1.

[0325] 107. The method of aspect 104, wherein said exosomes are 20 nm to 300 nm in size.

[0326] 108. The method of aspect 1 or aspect 2, wherein the extracellular vesicles are microvesicles and have a diameter between 100 and 1000 nm.

[0327] 109. The method of aspect 108, wherein said microvesicles have an irregular shape.

[0328] 110. The method of aspect 108, wherein said microvesicles express tissue factor.VABIO.002WO2 PCT APPLICATION

[0329] 111. The method of aspect 108, wherein the microvesicles express alpha3betal integrin.

[0330] 112. The method of aspect 108, wherein said microvesicles express markers found on the cell membrane of their cell of origin.

[0331] 113. The method of aspect 108, wherein said microvesicles express E-selectin.

[0332] 114. The method of aspect 1 or aspect 2, wherein said extracellular vesicles are apoptotic bodies.

[0333] 115. The method of aspect 114, wherein said apoptotic bodies possess phosphatidylserine on their membranes.

[0334] 116. The method of aspect 114, wherein said apoptotic bodies possess an irregular shape,

[0335] 117. The method of aspect 114, wherein said apoptotic bodies are 50-5000 nm in size.

[0336] 118. The method of aspect 114, wherein said apoptotic bodies are bound to histones.

[0337] 119. The method of aspect 1 or aspect 2, wherein said extracellular vesicles are administered together with a regenerative cell population.

[0338] 120. The method of aspect 1 or aspect 2, wherein conditioned media is used together or as a substitute for extracellular vesicles.

[0339] 121. The method of aspect 1 or aspect 2, wherein said regenerative cell population, alone, or together with conditioned media, and / or extracellular vesicles, is loaded into a suppository, or suppository-type device.

[0340] 122. The method of aspect 121, wherein said regenerative cell population is a population of pluripotent stem cells.

[0341] 123. The method of aspect 121, wherein said regenerative cell population is a population of multipotent stem cells.

[0342] 124. The method of aspect 121, wherein said regenerative cell population is a population of progenitor cells.

[0343] 125. The method of aspect 122, wherein said pluripotent stem cells are induced pluripotent stem cells.VABIO.002WO2 PCT APPLICATION

[0344] 126. The method of aspect 122, wherein said pluripotent stem cells are somatic cell nuclear transfer-derived pluripotent stem cells.

[0345] 127. The method of aspect 122, wherein said pluripotent stem cells are parthenogenesis-derived stem cells.

[0346] 128. The method of aspect 121, wherein said regenerative cells are hematopoietic stem cells

[0347] 129. The method of aspect 121, wherein said regenerative cells are mesenchymal stem cells and / or mesenchymal stromal cells.

[0348] 130. The method of aspect 121, wherein said regenerative cells are very small embryonic-like stem cells,

[0349] 131. The method of aspect 121, wherein said regenerative cells are mesenchymal stem cells and / or mesenchymal stromal cells.

[0350] The detailed description set forth herein is intended as a description of various configurations of the subject technology and is not intended to represent the only configurations in which the subject technology may be practiced. The detailed description provides specific details to facilitate an understanding of the subject technology. It will be apparent to those skilled in the art that the subject technology may be practiced without these specific details.

Claims

1. VABIO.002WO2 PCT APPLICATION2.WHAT IS CLAIMED IS:

1. A method of treating a subject having a disease, disorder, or condition, the method comprising:4.administering to the subject a composition comprising at least one primed stem cell, at least one primed extracellular vesicle, or a combination thereof,5.wherein the at least one primed stem cell underwent an at least one priming step m a media, the priming step comprising at least one of: inducing angiogenesis, administering a hormone, administering a pro-inflammatory molecule, culturing in 3D culture, or any combination thereof,6.wherein the at least one extracellular vesicle is derived from the stem cell or isolated from the cell-culture media following the at least one priming step.

2. The method of claim 1, wherein the disease, disorder, or condition comprises at least one of: a gender-specific disease, disorder, or condition, genitourinary syndrome of menopause, wound healing, scar treatment, loss of cell elasticity7, tissue damage, tissue dryness, tissue irritation, poor circulation, inflammation, infection, or any combination thereof.

3. The method of claim 2, wherein the disease, disorder, or condition is female¬ specific.

4. The method of claim 1, wherein angiogenesis is induced by the stem cell undergoing hypoxia.

5. The method of claim 1, wherein the pro-inflammatory molecule is a cytokine, preferably wherein the cytokine is IL-6.

6. The method of claim 1, wherein the hormone is estrogen.

7. A method of treating a subject having a disease, disorder, or condition associated with the female genitourinary tract, the method comprising:13.administering to the subject a biocompatible suppository formulated for dissolving inside the vaginal cavity of the subject, the suppository comprising a therapeutic composition, the therapeutic composition comprising at least one primed stem cell, at least one extracellular vesicle, or a combination thereof,14.wherein the at least one primed stem cell underwent an at least one priming step m a cell culture medium, the priming step comprising at least one of: inducing angiogenesis, VABIO.002WO2 PCT APPLICATION15.administering a hormone, administering a pro-inflammatory molecule, culturing in 3D culture, or any combination thereof,16.wherein the at least one extracellular vesicle is derived from the stem cell or isolated from the cell culture medium following the at least one priming step,17.wherein the at least one primed stem cell is of autologous, allogenic, or xenogeneic origin.

8. The method of claim 7, wherein the disease, disorder, or condition is selected from the group consisting of genitourinary inflammation, damage, dryness, irritation, degeneration, genitourinary syndrome of menopause, postpartum vaginal damage (PVD) Mayer-Rokita.nsky- Kiister-Hauser syndrome (MRKHS) following vaginoplasty, pelvic organ prolapse (POP), pelvic floor dysfunction, vaginitis, UTIs, female infertility caused by an immune imbalance in the genital urinary7tract, or any combination thereof.

9. The method of any one of the preceding claims, wherein the composition further comprises an antioxidant,10. The method of claim 9, wherein the antioxidant is selected from the group consisting of: ascorbic acid, alpha tocopherol, rutin, quercetin, hesperedin, lycopene, resveratrol, tetrahydrocur cumin, rosmarinic acid, Ellagic acid, chlorogemc acid, oleuropein, alpha-lipoic acid, glutathione, polyphenols, pycnogenol, and a derivative thereof.

11. The method of any one of the preceding claims, wherein the at least one primed stem cell is a committed progenitor cell, differentiated cell, or any combination thereof.

12. The method of claim 11, wherein the primed stem cell is an embryonic stem cell, cord blood stem cell, placental stem cell, bone marrow stem cell, amniotic fluid stem cell, neuronal stem cell, circulating peripheral blood stem cell, mesenchymal stem cell, mesenchymal stromal cell, germinal stem cell, adipose tissue derived stem cell, exfoliated teeth derived stem cell, hair follicle stem cell, dermal stem cell, partlienogenically derived stem cell, reprogrammed stem cell, side population stem cell, or any combination thereof.

13. The method of any one of the preceding claims, wherein the at least one primed stem cell is the mesenchymal stromal cell or an induced pluripotent stem cell (iPSC).

14. The method of any one of the preceding claims, wherein the at least one primed extracellular vesicle is derived from a mesenchymal stromal cell.VABIO.002WO2 PCT APPLICATION15. The method of claim 14, wherein the at least one primed extracellular vesicle is derived from a human umbilical mesenchymal stromal cell.

16. The method of any one of the preceding claims, wherein the at least one primed extracellular vesicle is isolated from animal blood.

17. The method of claim 16, wherein the blood is from a young animal.

18. The method of claim 17, wherein the blood is of a young sheep.

19. The method of any one of the preceding claims, wherein the at least one extracellular vesicle is between about 30 nm and about 150 nm m diameter.

20. The method of any one of the preceding claims, wherein the at least one primed stem cell, the at least one extracellular vesicle, or both, comprise a tetraspanin molecule.

21. The method of claim 20, wherein said tetraspanin molecule is a CD9, CD63, CD151, Tspan9, or Tspan32,22. The method of any one of the preceding claims, wherein the at least one primed stem cell, the at least one extracellular vesicle, or both, comprise at least one of a heat shock protein, a stress-associated protein, a growth factor receptor, or any combination thereof.

23. The method of claim 22, wherein the heat shock protein is selected calreticulin, hsp20, or hsp90.

24. The method of claim 22 or 23, wherein the stress-associated protein is phosphatidyl serine.

25. The method of any one of claims 22-24, wherein the growth factor receptor is IL-3 receptor, VEGF receptor, CXCL12, or c-met.

26. The method of any one of the preceding claims, wherein the biocompatible suppositoiy comprises chitosan HC1, cross-linking agent, collagen type I, carboxymethyl cellulose and DNase I27. The method of claim 21, wherein the weight ratio of chitosan HQ, cross-linking agent’s collagen type I, and carboxymethyl cellulose is about 40:30:30.

28. The method of any one of the preceding claims, wherein the biocompatible suppository’ also comprises a mucolytic enzyme.

29. The method of claim 23, wherein the mucolytic enzyme is DNase 1.

30. A biocompatible suppository formulated for dissolving inside the vaginal cavity of a subject, the suppository comprising a therapeutic composition with use against a disease,VABIO.002WO2 PCT APPLICATION41.disorder, or condition associated with the female genitourinary tract, wherein the therapeutic composition comprises at least one of:42.at least one stem cell; and43.at least one extracellular vesicle,44.wherein the at least one extracellular vesicle is derived from the stem cell or isolated from the biological fluid,45.wherein the at least one stem cell is of autologous, allogenic, or xenogeneic origin, and46.wherein the composition is primed to increase efficacy prior to being added to the suppository.47.31, The biocompatible suppository7of claim 25, wherein the subject is mammalian.

32. A composition with use against a disease, disorder, or condition associated with the female genitourinary tract, the composition comprising at least one of:48.at least one stem cell; and49.at least one extracellular vesicle,50.wherein the at least one extracellular vesicle is derived from the stem cell or isolated from the biological fluid,51.wherein the at least one stem cell is of autologous, allogenic, or xenogeneic origin, wherein the stem cell is primed to increase efficacy.

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