Therapeutic use of fibroblasts for the treatment of psoriasis

Abstract

Embodiments of the disclosure include methods and compositions for the treatment of psoriasis. In specific embodiments, fibroblasts, and / or fibroblast-derived materials, are utilized in at least some cases to affect the immune response of the individual in order to reduce the pathogenic levels of immune cells or cytokines and / or chemokines excreted by those immune cells.

Description

THERAPEUTIC USE OF FIBROBLASTS FOR THE TREATMENT OF PSORIASISCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application Serial No. 63 / 512,935, filed July 11, 2023, which is incorporated by reference herein in its entirety.BACKGROUNDI. Technical Field

[0002] This disclosure relates at least to the fields of cell biology, molecular biology, immunology, and medicine.II. Background

[0003] Fibroblasts are no longer considered as mere structural components of organs but as dynamic participants in immune processes. Fibroblasts produce an environment that influences T regulatory cell migration, proliferation, and activity to ensure immunotol erance

[0001] .

[0004] Psoriasis is a complex and chronic autoimmune inflammatory disease that afflicts approximately 2% of the world population and affects primarily the skin, nails, and joints. Psoriasis has a profound impact on quality of life, leading to an increase in the rate of anxiety and depress! on[2]. The disease is also associated with a significant number of comorbidities such as arthritis, cardiometabolic disease, diabetes mellitus, obesity, non-alcoholic fatty liver disease, inflammatory bowel disease, and depression[3-5],

[0005] Use of approved therapies utilizing small molecule inhibitors and humanized monoclonal antibodies to deplete and or modulate the immune system carry significant short and long-term side effects that can have a significant impact on quality of life. Using single cell fibroblasts, fibroblast spheroids, exosomes, lysates, apoptotic cells and or fibroblast- derived products which are natural immune modulators, achieve the same or better results in modulating the immune system, controlling the expansion of pathogenic immune cells, and their expression of pathogenic levels of cytokines, chemokines, and growth factors (e.g., see Liu et al. Adv Sci (Weinh). 2021 Aug; 8(15): 2004433, incorporated by reference herein in itsentirety) without the adverse side effect noted in currently approved therapies for psoriasis[6- 8].SUMMARY

[0006] Embodiments of the present disclosure include methods and compositions related to treatment or prevention of autoimmune inflammatory diseases, including an immune- mediated genetic skin disease, such as psoriasis. Individuals that are treated with methods and compositions of the disclosure include those that have one or more symptoms of psoriasis. Individuals that are prevented from having psoriasis, or in which the onset is delayed and / or the severity is reduced, include those at risk for psoriasis. Factors that predispose an individual to be at risk include extrinsic and intrinsic risk factors. Extrinsic risk factors include at least mechanical stress, air pollution, drugs, vaccination, infection, smoking, alcohol, or a combination thereof. Intrinsic risk factors include metabolic syndrome, obesity, diabetes mellitus, dyslipidemia, hypertension, mental stress, or a combination thereof. An individual may be genetically predisposed to psoriasis, including if they have a defect in the Psoriasis susceptibility 1 (PSORS1) gene, and the major susceptibility locus is HLA-Cw6.

[0007] Embodiments of the disclosure encompass compositions, methods, and systems for utilizing fibroblasts (including single cell fibroblasts), fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from fibroblast culture, any other fibroblast-derived material, and / or the like.

[0008] The fibroblasts and / or fibroblast-derived material(s) may be given to an individual that has psoriasis, or that might have psoriasis, or that is at risk for psoriasis for any reason, by any suitable route of delivery to the individual. In specific embodiments, the fibroblasts and / or fibroblast-derived material(s) may be delivered to the individual topically, subcutaneously, and / or intravenously, as examples only.

[0009] In some embodiments, the fibroblasts are modified ex vivo prior to delivery to the individual. In some embodiments, following delivery of the fibroblasts and / or fibroblast- derived material(s) to the individual, the resident fibroblast cells’ activities within the skin become normalized for regeneration of their function[9]. In certain embodiments, the fibroblasts are activated by one or more activating agents prior to use. In certain embodiments, the fibroblasts from which the fibroblast-derived materials are derived are activated by one or more activating agents prior to the use of the fibroblast-derived materials.

[0010] In particular embodiments, the fibroblasts are genetically modified. One or more endogenous genes in the fibroblasts may be modified, and / or the fibroblasts may harbor one ormore exogenously provided genes. Any gene modification system may be utilized, such as CRISPR / CAS systems. In specific cases, the gene modification system may be used to add, delete, or modify one or more fibroblast cell surface markers, e.g., to target the cells to a specific organ or tissue of interestflO], In some embodiments, the modified gene can be one or more of (1) a gene for a cytokine, such as, but not limited to, IL IRA, IL-4, IL-6, IL- 10, IL- 11, IL-13, TGFb, and / or the like; (2) a gene for soluble receptor, such as, but not limited to soluble TNF receptor p55, soluble TNF receptor p75, soluble IL-1 receptor type 2, membranebound IL-1 receptor type 2, IL- 18 binding protein, and / or the like; (3) a gene for a mediator such as, but not limited to, prostaglandin E2, and / or the like; (4) a gene for a cell surface receptor, such as, but not limited to PD1, CTLA4, TIM3, LAG3, TIGIT, CD96, BTLA, VISTA, and / or the like; and / or (5) other genes, such as, but not limited Arginase, Prostaglandin E2, Indoleamin 2,3-dioxygenase, Nitric oxide, and / or the like; and / or the like.

[0011] In particular embodiments, the disclosure concerns the interaction of fibroblasts with a second type of cells and / or certain agent(s) and includes modification(s) to the fibroblasts and / or second type of cells as a result of the interaction. In specific embodiments, the disclosure includes compositions, methods, and systems in which fibroblasts are modified upon exposure to certain cells and / or one or a combination of certain agent(s), including nucleic acids, cytokines, chemokines, or growth factors. In certain embodiments, delivery of these modified or unmodified fibroblasts to the individual (e.g., into the skin) can regenerate, repair, and / or reprogram the resident over-active functional fibroblasts, keratinocytes epithelial cells, and / or resident immune cells. In specific embodiments, this results in the initiation of the foundation for new tissue growth.

[0012] In certain embodiments, the regeneration of these fibroblasts, keratinocytes epithelial cells, and / or resident immune cells in the individual can be either through one or more possible steps, such as:

[0013] (1) The differentiation of the modified (e.g., such as by one or more activating agents, gene modification, or both) or unmodified fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from fibroblast, any other fibroblast-derived materials, and / or the like. The delivery to the individual may be intravenously, subcutaneously, or topically, including by injection.

[0014] (2) Epigenetic reprogramming of senescent or pathogenic fibroblasts, senescent or pathogenic keratinocytes, and / or senescent or pathogenic epithelial cells in the individual through contact with the delivered modified or unmodified fibroblasts, fibroblast spheroids,fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from fibroblast, any other fibroblast-derived materials, and / or the like.

[0011] ,

[0015] (3) Reprogramming or activating pathogenic fibroblasts, over-activated keratinocytes, skin resident immune cells by one or more types of products excreted from the modified or unmodified fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from fibroblast, or any other fibroblast- derived materials, and / or the like in the skin tissue microenvironment (e.g., the area of the skin affected by psoriasis).

[0016] (4) Recruitment, activation, and differentiation of local stem cells into active fibroblasts, keratinocytes, and or epithelial cells through the direct / indirect action of the injected modified / unmodified single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from fibroblast, and / or fibroblast-derived materials from the modified / unmodified fibroblasts.

[0017] (5) Modulation of the immune system through single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from fibroblasts, and / or fibroblast-derived materials to control the expansion and localization of pathogenic immune cells, and their expression of pathogenic level of any one or more cytokines, one or more chemokines and / or one or more growth factors associated with inflammation associated with psoriasis. These cells include T cells, dendritic cells, macrophages, B cells, and neutrophils, expressing pathogenic levels of one or more of IL-23, IL-1, IL-10, IL-22, IL-17, IL-12, TNF, INF- y, TGF- 1, as examples.

[0018] The regeneration of these fibroblasts, keratinocytes, epithelial cells, and / or resident immune cells may occur by one or more other mechanisms.

[0019] In certain embodiments, methods and compositions herein allow for the reduction of the pathogenic levels of immune cells and their impact on the skin cells in psoriasis. Such cells can be referred to herein as “psoriasis-associated immune cells.” Such cells can be, but are not limited to pathogenic T cells, pathogenic dendritic cells, pathogenic macrophages, and / or pathogenic neutrophils, and / or the like. Such cells can express pathogenic levels of IL- 23, IL-1, IL- 10, IL-22, IL- 17, IL- 12, TNF, INF- y, TGF- 1, and / or the like. Reduction of inflammation and levels of pathogenic cytokines, pathogenic chemokines, and / or pathogenic growth factors will lead to reprogramming of the hyper-keratinized inflamed skin. In certain embodiments, the methods and compositions allow for normalizing the immune response in order to reduce the pathogenic levels of immune cells or cytokines, growth factors, and / or chemokines excreted by those cells.

[0020] In certain embodiments, the disclosure concerns compositions, methods, and systems in which certain cells are modified upon exposure to fibroblasts and / or certain agent(s) excreted from fibroblasts. In certain embodiments, the interaction of fibroblasts with one or more other types of cells (and optionally, that interaction also includes one or more certain agent(s)) results in modification of the fibroblasts and / or the other type of cells. In certain embodiments, the other types of cells include at least immune cells.

[0021] In certain embodiments, the methods of the disclosure occur ex vivo, such as in a culture and can also occur in vivo. In particular cases, the methods occur by the hand of a person and do not encompass ordinary or random occurrences in a body. The methods of the disclosure are non-natural, in particular aspects. In certain embodiments, the concentrations of cells used in a method of exposing one type of cells to another type of cells does not occur in nature and does not happen randomly in nature. In certain embodiments, the concentration of one or more agents used in a method of exposing one or more agents to one or more types of cells does not occur in nature and does not happen randomly in nature. The modification of any type of cells encompassed by the disclosure that occurs ex vivo or in vitro does not occur in vivo naturally in the same manner. In such embodiment, tissue biopsy from the donor is used to isolate, characterize, if necessary, activate, expand, and reintroduce back into the donor one or a combination of the cells for the purpose of modulating the immune system, reprogramming local keratinocytes, fibroblasts, and or epithelial cells.

[0022] The disclosure encompasses therapeutic uses of cells, including epithelial cells, keratinocytes, epithelial cells, dendritic cells, macrophages, B lymphocytes, myoid cells, endothelial cells, fibroblasts, immune cells, and mixtures thereof. In at least some cases, the single cell fibroblasts, fibroblast spheroids, or fibroblast-derived materials have been modified prior to their exposure to the immune cells and other cells found in the blood and skin tissue, such as chemically, physically, and / or epigenetically activated, and / or exposed to conditions that are not normally found in the body, and in other cases, immune cells, or their derivatives, have been modified, such as activated, prior to their exposure to the fibroblasts and / or fibroblast-derived materials.

[0023] Embodiments of the disclosure provide means of utilizing fibroblasts as allogeneic, autologous, xenogeneic, and / or syngeneic therapeutic cells through modification of culture conditions. In one embodiment of the disclosure, fibroblasts are extracted from sources with lower immunogenicity (e.g., placental fibroblasts, omental tissue-derived fibroblasts, cord blood-derived fibroblasts, etc. .

[0024] In one embodiment of the disclosure, single cell fibroblasts and fibroblast spheroids are cultured in vitro for preserving the viability and proliferative ability of fibroblasts. The disclosure provides for the modification of known culture techniques to decrease the recognition of fibroblasts by the recipient’s immune system. In one embodiment, fibroblasts, and / or fibroblast spheroids (for example) may be cultured in conditions that lack xenogeneic components, such as xenogeneic-free medium; in some cases, for example, the media may be free of fetal calf serum. In certain embodiments, the disclosure encompasses the substitution of fetal calf serum with one or more other agents, such as those that facilitate the reduction of immunogenicity of fibroblasts, for example, human platelet-rich plasma, platelet lysate, umbilical cord blood serum, autologous serum, and / or one or more defined cytokines, such as one or a combination of fibroblast growth factor, epidermal growth factor, leukemia inhibitory factor, insulin-like growth factor, angiopoietin, and vascular endothelial growth factor.

[0025] In one embodiment of the disclosure, effective amounts of single cell fibroblasts, fibroblast spheroids, or fibroblast-derived materials as prepared in methods encompassed by the disclosure are administered to an individual for therapy or prevention of one or more medical conditions. In certain embodiments, the fibroblasts are administered to improve keratinocyte, epithelial, fibroblast, and / or resident immune cell responsiveness, modulation of immune cell activity, and cell differentiation.

[0026] Embodiments of the disclosure provide methods for the co-administration of universal donor single cell fibroblasts, fibroblast spheroids, or fibroblast-derived materials with one or more agents that modulate the immune cell activity in the site of inflammation. In a certain embodiment of the disclosure, methods are provided for the co-administration of universal donor single cell fibroblasts, fibroblast spheroids, or fibroblast-derived materials with one or a combination of growth factors, chemokines, and / or cytokines. In one embodiment of the disclosure, universal donor fibroblasts derived from fibroblasts that have been treated under conditions to reduce immunogenicity are utilized to stimulate vascular endothelial growth factor (VEGF) production by the introduced modified / unmodified fibroblasts or from endogenous cells under the regulatory control of nerve growth factor (NGF) of the individual thereby leading to tissue regeneration and revascularization of the damaged skin.

[0027] Embodiments of the disclosure provide methods for reducing the immunogenicity of particular types of fibroblasts. Single cell fibroblasts, fibroblast spheroids, exosomes, lysates, apoptotic cells, or fibroblast-derived materials may be derived from various tissues or organs, including, but not limited to, skin, heart, blood vessels, bone marrow, skeletal muscle, liver, pancreas, brain, foreskin, which can be obtained by biopsy (where appropriate) or uponautopsy. In some aspects, the cells comprise fibroblasts, which can be of fetal, neonatal, adult origin, or a combination thereof.

[0028] Single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials or products for use in any methods of the disclosure may be exposed to certain medium component(s), in certain embodiments.

[0029] In some embodiments, one may monitor the levels of particular immune cells and / or one or more agents expressed by the cells. This may occur prior to treatment, during treatment, and / or following treatment. Levels of T cells, dendritic cells, B cells, macrophages, and / or neutrophils may be monitored by Complete Blood Count and White Blood Count, and tissue levels of T cells, dendritic cells, macrophages, and IL-23, IL-1, IL-10, IL-22, IL-17, IL-12, TNF, INF- y, TGF- pi may be monitored using ELISA and / or bead-based immunoassays, for example.

[0030] Embodiments of the disclosure encompass an in vivo method of treating psoriasis using a topical, intravenous, or subcutaneous injection of single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from fibroblast, and / or fibroblast-derived materials, such as to modulate the immune system and to reduce the proliferation of psoriasis-associated immune cells. Psoriasis- associated immune cells can be pathogenic T cells, pathogenic dendritic cells, pathogenic macrophages, and / or pathogenic neutrophils, and / or the like that express pathogenic levels of IL-23, IL-1, IL-10, IL-22, IL-17, IL-12, TNF, INF- y, TGF-pi, and / or the like. The fibroblast may be autologous, allogeneic, xenogeneic, or syngeneic with respect to the individual, and in some cases, a combination may be utilized.

[0031] Particular embodiments include introduction into an individual of single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from fibroblast, and / or fibroblast-derived materials, or these products wherein the fibroblasts may be activated with one or more agents. In some cases, the single cell fibroblasts, fibroblast spheroids, or fibroblast-derived materials plus one or more types of non-pathogenic immune cells or derivative agents thereof are delivered into the bloodstream or skin tissue to modulate the proliferation of pathogenic immune cells therein. When the fibroblasts, fibroblast spheroids, or fibroblast-derived materials come into the presence of or into contact with non-pathogenic immune cells, this may or may not occur using a co-culture method prior to administration. Examples of non-pathogenic immune cells may include T cells, dendritic cells, macrophages, neutrophils, NK cells, or a combination thereof. In certaincases, the non-pathogenic immune cells do not express pathogenic levels of IL-23, IL-1, IL- 10, IL-22, IL- 17, IL- 12, TNF, INF- y, and / or TGF- pi.

[0032] Some embodiments include the introduction of single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from fibroblast, and / or fibroblast-derived materials, wherein these may be activated with one or more agents or conditions. In some cases, the single cell fibroblasts, fibroblast spheroids, or fibroblast-derived materials plus one or more types of immune cells and / or derivative agents are provided into the blood-stream and / or skin tissue of the individual to modulate the expression of pathogenic levels of IL-23, IL-1, IL-10, IL-22, IL-17, IL-12, TNF, INF- y, TGF-pi.

[0033] Particular embodiments include the introduction of single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials, single cell fibroblasts, fibroblast spheroids, or fibroblast-derived materials or products activated with one or more agents or conditions. In some cases, the single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from the fibroblast, and / or fibroblast-derived materials are administered to an individual with one or more types of immune cells and / or one or more derivative agents. The single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from the fibroblast, and / or fibroblast-derived materials plus one or more types of immune cells and / or one or more derivative agents can be introduced into the blood-stream and / or skin tissue of the individual. The administration can enable the epigenetic reprogramming of resident fibroblasts in the skin, resident keratinocytes in the skin, and / or resident epithelial cells in the skin of the individual. The one or more types of immune cells and / or one or more derivative agents can be administered to the individual simultaneously in a single or separate composition as the single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials, single cell fibroblasts, fibroblast spheroids, or fibroblast-derived materials, or can be administered at a different time in a different composition. In some embodiments, the one or more types of immune cells and / or one or more derivative agents can be administered in one composition before or after administration of the single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials, single cell fibroblasts, fibroblast spheroids, or fibroblast-derived materials in a different composition. The different compositions can be administered in the same way or different ways. For example,one composition can be administered into the blood stream of an individual (e.g., intravenously, subcutaneously, etc.) and the other composition can be administered onto or into or near the skin of the individual (e.g., topically, subcutaneously, etc.).

[0034] Introduction of single cell fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, conditioned media from fibroblast, and / or fibroblast-derived materials or products, may be activated with one or more agents. In some cases, the single cell fibroblasts, fibroblast spheroids, or fibroblast-derived materials plus one or more types of immune cells and / or one or more derivative agents thereof are introduced into the blood- stream and / or skin tissue of the individual to enable the activation and migration of localized stem cell niches to replace the pathogenetic keratinocytes, pathogenetic fibroblasts, and / or pathogenetic epithelial cells.

[0035] In some embodiments, there is the use of one or more adjuvants in combination with single cell fibroblasts, fibroblast spheroids, and / or fibroblast-derived materials to activate the immune system. These adjuvants could be chemical, viral, or bacterial product based.

[0036] The foregoing has outlined rather broadly the features and technical advantages of the present invention so that the detailed description of the invention that follows may be better understood. Additional features and advantages of the invention will be described hereinafter, which form the subject of the claims of the invention. It should be appreciated by those skilled in the art that the conception and certain embodiment disclosed may be readily utilized as a basis for modifying or designing other structures for carrying out the same purposes of the present invention. It should also be realized by those skilled in the art that such equivalent constructions do not depart from the spirit and scope of the invention as set forth in the appended claims. The novel features that are believed to be characteristic of the invention, both its organization and method of operation, together with further objects and advantages, will be better understood from the following description when considered in connection with the accompanying figures. It is to be expressly understood, however, that each of the figures is provided for the purpose of illustration and description only and is not intended as a definition of the limits of the present invention.BRIEF DESCRIPTION OF THE DRAWINGS

[0037] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be betterunderstood by reference to one or more of these drawings in combination with the detailed description of certain embodiments presented herein.

[0038] FIG. 1. Clinical score for erythema (a scale from 0 to 4, 0-none; 1 -slight; 2- moderate; 3-marked; 4-very marked) for experiments described in the Examples.

[0039] FIG. 2. Clinical score for scaling (a scale from 0 to 4, 0-none; 1 -slight; 2-moderate; 3-marked; 4-very marked) for experiments described in the Examples.

[0040] FIG. 3. Clinical score for thickness (a scale from 0 to 4, 0-none; 1 -slight; 2- moderate; 3-marked; 4-very marked) for experiments described in the Examples.

[0041] FIG. 4. Average of all clinical scores for experiments described in the Examples. A single intravenous administration of fibroblasts on day 4 significantly alleviated imiquimod (IMQ)-induced psoriasis from day 6 to day 8, with a remarkable 35% decrease in the average clinical score (from 3.4 to 2.2) on day 8. In mice with moderate-to-severe psoriasis, multiple administrations of HDF spheroids on day 7, day 10, and day 13 led to a notable reduction in the average clinical scores by 18% on day 10, 26% on day 13, and 36% on day 15.

[0042] FIG. 5. Skin images of the control and the fibroblast-treated mice. The control group intravenous injection of vehicle (Plasma-Lyte A) and the fibroblast treatment group intravenous injection were carried out on day 4 for the single administration study and days 7, 10, and 13 for the multiple administration study. By terminations (day 8 and day 15), the psoriasis clinical indications were significantly reduced in the treatment group compared to the control group.

[0043] FIG. 6. Skin H&E images showed a substantial 4.8-fold increase in epidermal thickness within the lesions in psoriatic mice compared to healthy controls. A single administration of fibroblasts significantly mitigated this effect, resulting in a remarkable reduction in epidermal thickness (from 131.94 ± 3.52 pm to 74 ± 4.32 pm).

[0044] FIG. 7. Skin immunohistochemical staining for CD3 (lymphocytes) and F4 / 80 (macrophages) revealed treatment with fibroblasts resulted in a down-regulation of immune cell infiltration within the psoriatic lesions.

[0045] FIG. 8. Skin immunofluorescent staining for Ki67 (cell proliferation) showed fibroblasts inhibited the proliferation of keratinocytes within the epidermis, resulting in decreased epidermal thickness in fibroblast-treated psoriatic mice.

[0046] FIG. 9. Fibroblasts significantly attenuated the IMQ-induced increase in spleen weight and length.

[0047] FIG. 10. Fibroblasts improved red blood cells, hemoglobin, and platelets in mice with IMQ-induced psoriasis. IMQ-induced immune cell activation, evident by the increase in neutrophils and monocytes in the peripheral blood, was significantly inhibited by fibroblasts.DETAILED DESCRIPTIONI. Examples of Definitions

[0048] In keeping with long-standing patent law convention, the words “a” and “an” when used in the present specification in concert with the word comprising, including the claims, denote “one or more.” Some embodiments of the disclosure may consist of or consist essentially of one or more elements, method steps, and / or methods of the disclosure. It is contemplated that any method or composition described herein can be implemented with respect to any other method or composition described herein.

[0049] The phrase “and / or” means “and” or “or”. To illustrate, A, B, and / or C includes: A alone, B alone, C alone, a combination of A and B, a combination of A and C, a combination of B and C, or a combination of A, B, and C. In other words, “and / or” operates as an inclusive or.

[0050] As used herein, the term “about” or “approximately” refers to a quantity, level, value, number, frequency, percentage, dimension, size, amount, weight, or length that varies by as much as 30, 25, 20, 25, 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 % to a reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight or length. In particular embodiments, the terms “about” or “approximately” when preceding a numerical value indicates the value plus or minus a range of 15%, 10%, 5%, or 1%. With respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5- fold, and more preferably within 2-fold, of a value. Unless otherwise stated, the term 'about' means within an acceptable error range for the particular value.

[0051] 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’ 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 inthe disclosure for the listed elements. Thus, the phrase “consisting essentially of’ indicates that the listed elements are required or mandatory, but that no other elements are optional and may or may not be present depending upon whether or not they affect the activity or action of the listed elements.

[0052] The term "comprising," which is synonymous with "including," "containing," or "characterized by," is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. The phrase "consisting of excludes any element, step, or ingredient not specified. The phrase "consisting essentially of limits the scope of described subject matter to the specified materials or steps and those that do not materially affect its basic and novel characteristics. It is contemplated that embodiments described in the context of the term "comprising" may also be implemented in the context of the term "consisting of or "consisting essentially of."

[0053] Reference throughout this specification to “one embodiment,” “an embodiment,” “a particular embodiment,” “a related embodiment,” “a certain embodiment,” “an additional embodiment,” or “a further embodiment” or combinations thereof means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. Thus, the appearances of the foregoing phrases in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0054] As used herein, the term "activated fibroblasts" refers to any kind of fibroblasts treated with one or more stimuli or agent(s) capable of inducing one or more alterations in the fibroblast: metabolic, immunological, epigenetic, growth factor-secreting, surface marker expression, and production and excretion of microvesicles. Examples of agents include at least ADP, CXCL4, PDGF, TGFp, IL-I, TNFa, ROS, NO, AMP, IL-17, IL22, dsRNA 1.25D3, IL- 8, CXCLI-8, Activin A, P-defensins, cathelicidin, E-selectin, P-selectin, KGF, VEGF, IGF, IL- 10, IL-4, and IL-13.

[0055] The term “derivative agents of immune cells” refers to any agent associated with any immune cell. For example, any agent produced by any immune cell. For example, cytokines and / or chemokines and / or growth factors and / or the like.

[0056] As used herein, the term “activated immune cells” refers to immune cells treated with one or more stimuli capable of inducing one or more alterations in the cell: metabolic, immunological, epigenetic, or growth factor secreting. Examples of agents include at leastADP, CXCL4, PDGF, TGFp, IL-I, TNFa, ROS, NO, AMP, IL-17, IL22, dsRNA 1.25D3, IL- 8, CXCLI-8, Activin A, P-defensins, cathelicidin, E-selectin, P-selectin, KGF, VEGF, IGF, IL- 10, IL-4, and IL-13.

[0057] The term "administered" or "administering", as used herein, refers to any method of providing a composition to an individual such that the composition has its intended effect on the individual. For example, one method of administering is by a direct mechanism such as local tissue administration, intravenous administration, oral ingestion, transdermal patch, topical, inhalation, suppository, etc. Another example includes an indirect mechanism using a medical device such as, but not limited to, a catheter, applicator gun, syringe, etc.

[0058] As used herein, “allogeneic” refers to tissues or cells from another body that, in a natural setting, are immunologically incompatible or capable of being immunologically incompatible, although from one or more individuals of the same species.

[0059] As used herein, “autologous” refers to tissues or cells that are derived or transferred from the same individual's body (i.e., autologous blood donation; an autologous bone marrow transplant).

[0060] As used herein, “agent” refers to nucleic acids, cytokines, chemokines, transcription factors, epigenetics factors, growth factors, or hormones.

[0061] As used herein, “xenogeneic” refers to tissues or cells from a species different from the patient.

[0062] Cell culture" may be an artificial in vitro system containing viable cells, whether quiescent, senescent or (actively) dividing. In a cell culture, cells are grown and maintained at an appropriate temperature, typically a temperature of 37°C and under an atmosphere typically containing oxygen and CO2. Culture conditions may vary widely for each cell type though, and variation of conditions for a particular cell type can result in different phenotypes being expressed. The most commonly varied factor in culture systems is the growth medium. Growth media can vary in concentration of nutrients, growth factors, and the presence of other components. The growth factors used to supplement media are often derived from animal blood, such as calf serum.

[0063] The term “fibroblast-derived material” as used herein refers to fibroblast cell fragments, fibroblast conditioned media, exosomes secreted from fibroblasts, and / or fibroblast lysate and includes fibroblast-like cells; and / or extracellular vesicles including exosomes, microvesicles, apoptotic bodies or any other fragments or biologic components of fibroblast cells.

[0064] The term "fibroblast-like cell" refers to cells that share certain characteristics of fibroblasts, but exhibit surface markers, structural features, expression patterns, and epigenetic profiles similar to those of the cells they are closely associated with in a specific location. Examples include myofibroblasts, perineurial sheath cells, lymphoid organ dendritic cells, and villus fibroblasts in the intestine.

[0065] The term "individual", as used herein, refers to a human or animal that may or may not be housed in a medical facility and may be treated as an outpatient of a medical facility. The individual may be receiving one or more medical compositions via the internet. An individual may comprise any age of a human or non-human animal and therefore includes both adults and juveniles (z.e., children) and infants. It is not intended that the term "individual" connotes a need for medical treatment, therefore, an individual may voluntarily or involuntarily be part of experimentation, whether clinical or in support of basic science studies. The term “subject” or “individual” may be used interchangeably and refers to any organism or animal subject that is an object of a method or material, including mammals, e.g., humans, laboratory animals (e.g., primates, rats, mice, rabbits), livestock (e.g., cows, sheep, goats, pigs, turkeys, and chickens), household pets (e.g., dogs, cats, and rodents), horses, and transgenic non-human animals.

[0066] The terms "reduce," "inhibit," "diminish," "suppress," "decrease," "prevent" and grammatical equivalents (including "lower," "smaller," etc.) when in reference to the expression of any symptom in an untreated subject relative to a treated subject, mean that the quantity and / or magnitude of the symptoms in the treated subject is lower than in the untreated subject by any amount that is recognized as clinically relevant by any medically trained personnel. In one embodiment, the quantity and / or magnitude of the symptoms in the treated subject is at least 10% lower than, at least 25% lower than, at least 50% lower than, at least 75% lower than, and / or at least 90% lower than the quantity and / or magnitude of the symptoms in the untreated subject.

[0067] As used herein, the term "transplantation" refers to the process of taking living tissue or cells and implanting it in another part of the body or into another body. As used herein, the term "therapeutically effective amount" is the amount of a composition provided to an individual such that a wound is improved in severity and / or size (including diameter or area).

[0068] Treatment,” “treat,” or “treating” means a method of reducing the effects of a disease or condition. Treatment can also refer to a method of reducing the disease or condition itself rather than just the symptoms. The treatment can be any reduction from pre-treatment levels and can be but is not limited to the complete ablation of the disease, condition, or thesymptoms of the disease or condition. Therefore, in the disclosed methods, treatment” can refer to a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% reduction in the severity of an established disease or the disease progression, including reduction in the severity of at least one symptom of the disease. For example, a disclosed method for reducing the immunogenicity of cells is considered to be a treatment if there is a detectable reduction in the immunogenicity of cells when compared to pre-treatment levels in the same subject or control subjects. Thus, the reduction can be 10, 20, 30, 40, 50, 60, 70, 80, 90, 100%, or any amount of reduction in between as compared to native or control levels. It is understood and herein contemplated that “treatment” does not necessarily refer to a cure for the disease or condition, but an improvement in the outlook of a disease or condition. In certain embodiments, treatment refers to the lessening in severity or extent of at least one symptom and may alternatively or in addition refer to a delay in the onset of at least one symptom.

[0069] As used herein, “blow-mold” means a method of creating hollow products from thermoplastic material. As used herein, “injection-mold” means a method of creating solid plastic parts using precision molds and tooling.

[0070] Although the present disclosure and its advantages have been described in detail, it should be understood that various changes, substitutions, and alterations can be made herein without departing from the spirit and scope of the disclosure as defined by the appended claims. Moreover, the scope of the present application is not intended to be limited to the particular embodiments of the process, machine, manufacture, composition of matter, means, methods, and steps described in the specification. As one of ordinary skill in the art will readily appreciate from the disclosure of the present disclosure, processes, machines, manufacture, compositions of matter, means, methods, or steps presently existing or later to be developed that perform substantially the same function or achieve substantially the same result as the corresponding embodiments described herein may be utilized according to the present disclosure. Accordingly, the appended claims are intended to include within their scope such processes, machines, manufacture, compositions of matter, means, methods, or steps.

[0071] It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method or composition of the invention, and vice versa. Furthermore, compositions of the invention can be used to achieve methods of the invention.

[0072] Other objects, features and advantages of the present invention will become apparent in the following detailed description. It should be understood, however, that the detailed description and certain examples, while indicating certain embodiments of the invention, are given by way of illustration only, since various changes and modifications withinthe spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.II. Administration of Therapeutic Compositions

[0073] Particular embodiments of the disclosure concern therapeutic compositions for the treatment of psoriasis. The compositions may cure psoriasis of the individual or may reduce the severity of one symptom of psoriasis and / or delay the onset of psoriasis. In certain embodiments, the individual has symptoms such as, but not limited to, dry skin lesions, known as plaques, covered in scales. The plaques may be itchy sore, or both. In severe cases, the skin around the joints may crack and bleed. In certain embodiments, they are on the elbows, knees, scalp and / or lower back, but they may be anywhere on the body.

[0074] The psoriasis may be of any kind, including at least plaque psoriasis, scalp psoriasis, nail psoriasis, guttate psoriasis, inverse (flexural) psoriasis, pustular psoriasis, von Zumbusch psoriasis, palmoplantar pustulosis, erythrodermic psoriasis, and / or the like.

[0075] In some embodiments, the methods and compositions of the disclosure are utilized in addition to one or more other psoriasis treatments, such as Steroid creams, roflumilast, moisturizers, salicylic acid, coal tar, vitamin D-based cream or ointment, retinoid creams, calcineurin inhibitors, anthralin, and / or the like. In some embodiments, the individual in need of the methods and compositions of the disclosure has employed one or more of these other psoriasis treatments, such as to little or no effect.

[0076] The therapeutic agents of the disclosure may be administered by any suitable administration route. When more than one psoriasis therapy is utilized, the therapeutic agent of the disclosure may be administered by the same route of administration or by different routes of administration than the additional therapy. In some embodiments, the therapy is administered intravenously, topically, intramuscularly, subcutaneously, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally. In some embodiments, the antibiotic is administered intravenously, intramuscularly, subcutaneously, topically, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, intranasally, and / or the like. The appropriate dosage may be determined based on the type of disease to be treated, the severity and course of the disease, the clinical condition of the individual, the individual's clinical history and response to the treatment, and the discretion of the attending physician.

[0077] The treatments may include various “unit doses.” Unit dose is defined as containing a predetermined-quantity of the therapeutic composition. The quantity to be administered, and the particular route and formulation, are within the skill of determination of those in the clinical arts. A unit dose need not be administered as a single injection but may comprise continuous infusion over a set period of time. In some embodiments, a unit dose comprises a single administrable dose.

[0078] In some embodiments, a single dose of the therapy is administered. In some embodiments, multiple doses of the therapy are administered. In some embodiments, the therapy is administered at a dose of 100 cells up to 10 million cells. In some embodiments, the therapy is administered at a dose of at least, at most, or about 100-1 million, 100-100,000, 100- 10,000, 100-1000, 1000-10 million, 1000-1 million, 1000-100,000, 1000-10,000, 10,000-10 million, 10,000-1 million, 10,000-100,000, 100,000-10 million, 100,000-1 million, or 1 million -10 million cells, or products therefrom, or any range derivable therein.

[0079] The quantity to be administered, both according to the number of treatments and unit dose, depends on the treatment effect desired. An effective dose is understood to refer to an amount necessary to achieve a particular effect. In practice, in certain embodiments, it is contemplated that doses in the range from 100 cells-10 million cells, or products therefrom, can affect the protective capability of these agents. Thus, it is contemplated that doses include doses of about 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, and 200, 300, 400, 500, 1000 pg / kg, mg / kg, pg / day, or mg / day or any range derivable therein. Furthermore, such doses can be administered at multiple times during a day and / or on multiple days, weeks, or months.

[0080] Precise amounts of the therapeutic composition also depend on the judgment of the practitioner and may be tailored to each individual. Factors affecting dose include physical and clinical state of the patient, the route of administration, the intended goal of treatment (alleviation of symptoms versus cure) and the potency, stability and toxicity of the particular therapeutic substance or other therapies a subject may be undergoing.

[0081] It is also understood that uptake is species and organ / tissue-dependent. The applicable conversion factors and physiological assumptions to be made concerning uptake and concentration measurement are well-known and would permit those of skill in the art to convert one concentration measurement to another and make reasonable comparisons and conclusions regarding the doses, efficacies and results described herein.

[0082] In certain instances, it will be desirable to have multiple administrations of the composition, e.g., 2, 3, 4, 5, 6 or more administrations. The administrations can be at 1, 2, 3, 4, 5, 6, 7, 8, to 5, 6, 7, 8, 9, 10, 11, or 12-week intervals, including all ranges there between.

[0083] The phrases “pharmaceutically acceptable” or “pharmacologically acceptable” refer to molecular entities and compositions that do not produce an adverse, allergic, or other untoward reaction when administered to an animal or human. As used herein, “pharmaceutically acceptable carrier” includes any and all solvents, dispersion media, coatings, anti-bacterial and anti-fungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well-known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredients, its use in immunogenic and therapeutic compositions is contemplated. Supplementary active ingredients, such as other anti-infective agents and vaccines, can also be incorporated into the compositions.

[0084] The active compounds can be formulated for parenteral administration, e.g., formulated for injection via the intravenous, intramuscular, subcutaneous, or intraperitoneal routes. Typically, such compositions can be prepared as either liquid solutions or suspensions; solid forms suitable for use to prepare solutions or suspensions upon the addition of a liquid prior to injection can also be prepared; and the preparations can also be emulsified.

[0085] The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including, for example, aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form may be sterile and fluid to the extent that it may be easily injected. It also should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.

[0086] The proteinaceous compositions may be formulated into a neutral or salt form. Pharmaceutically acceptable salts, include the acid addition salts (formed with the free amino groups of the protein), which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine, and the like.

[0087] A pharmaceutical composition can include a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The properfluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various anti-bacterial and anti-fungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0088] Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent with various other ingredients enumerated above, as required, followed by filtered sterilization or an equivalent procedure. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle, which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques, which yield a powder of the active ingredient, plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0089] Administration of the compositions will typically be via any common route. This includes, but is not limited to topical and / or intravenous administration. In some embodiments, both topical and intravenous methods are utilized. For example, a first dose may be a bolus that is intravenous, followed by topical administration thereafter. Alternatively, administration may be by orthotopic, intradermal, subcutaneous, intramuscular, intraperitoneal, or intranasal administration. Such compositions would normally be administered as pharmaceutically acceptable compositions that include physiologically acceptable carriers, buffers, or other excipients.

[0090] Upon formulation, solutions are administered in a manner compatible with the dosage formulation and in such amount as is therapeutically or prophylactically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above.III. Cellular TherapiesA. Cells and Cell Culture

[0091] In certain embodiments, the fibroblasts may be derived from tissues comprising skin, heart, blood vessels, bone marrow, skeletal muscle, liver, pancreas, brain, adipose tissue,foreskin, placental, and / or umbilical cord. In certain embodiments, the fibroblasts are placental, fetal, neonatal, adult, or mixtures thereof.

[0092] In some embodiments, fibroblast cells may be cultured for at least between about 10 days and about 40 days, for at least between about 15 days and about 35 days, for at least between about 15 days and 21 days, such as for at least about 15, 16, 17, 18, 19 or 21 days. In some embodiments, the cells of the disclosure may be cultured for no longer than 60 days, no longer than 50 days, or no longer than 45 days. The cells may be cultured for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 days. The cells may be cultured in the presence of a liquid culture medium. Typically, the medium may comprise a basal medium formulation as known in the art. Many basal media formulations can be used to culture cells herein, including but not limited to Eagle's Minimum Essential Medium (MEM), Dulbecco's Modified Eagle's Medium (DMEM), alpha modified Minimum Essential Medium (a-MEM), Basal Medium Essential (BME), Iscove's Modified Dulbecco's Medium (IMDM), BGJb medium, F-12 Nutrient Mixture (Ham), Liebovitz L-15, DMEM / F-12, Essential Modified Eagle's Medium (EMEM), RPMI-1640, and modifications and / or combinations thereof. Compositions of the above basal media are generally known in the art, and it is within the skill of one in the art to modify or modulate concentrations of media and / or media supplements as necessary for the cells cultured. In some embodiments, a culture medium formulation may be explants medium (CEM), which is composed of IMDM supplemented with 10% fetal bovine serum (FBS), 100 U / ml penicillin G, 100 pg / ml streptomycin and 2 mmol / L L-glutamine. Other embodiments may employ further basal media formulations, such as those chosen from the ones above.

[0093] Any medium capable of supporting cells in vitro may be used to culture the cells. Media formulations that can support the growth of cells include, but are not limited to, Dulbecco's Modified Eagle's Medium (DMEM), alpha modified Minimal Essential Medium (aMEM), and Roswell Park Memorial Institute Media 1640 (RPMI Media 1640) and the like. Typically, up to 20% fetal bovine serum (FBS) or 1-20% horse serum is added to the above medium in order to support the growth of cells. A defined medium, however, can also be used if the growth factors, cytokines, and hormones necessary for culturing cells are provided at appropriate concentrations in the medium. Media useful in the methods of the disclosure may comprise one or more compounds of interest, including, but not limited to, antibiotics, mitogenic compounds, or differentiation compounds useful for the culturing of cells. The cells may be grown at temperatures between 27° C to 40° C, such as 31° C to 37° C, and may be in a humidified incubator. The carbon dioxide content may be maintained between 2% to 10%,and the oxygen content may be maintained between 1% and 22%. The disclosure, however, should in no way be construed to be limited to any one method of isolating and culturing cells. Rather, any method of isolating and culturing cells should be construed to be included in the present disclosure.

[0094] For use in the cell culture, media can be supplied with one or more further components. For example, additional supplements can be used to supply the cells with the necessary trace elements and substances for optimal growth and expansion. Such supplements include insulin, transferrin, selenium salts, and combinations thereof. These components can be included in a salt solution such as, but not limited to, Hanks' Balanced Salt Solution (HBSS) and Earle's Salt Solution. Further antioxidant supplements may be added, e.g., P- mercaptoethanol. While many media already contain amino acids, some amino acids may be supplemented later, e.g., L-glutamine, which is known to be less stable when in solution. A medium may be further supplied with antibiotic and / or antimycotic compounds, such as, typically, mixtures of penicillin and streptomycin, and / or other compounds, exemplified but not limited to, amphotericin, ampicillin, gentamicin, bleomycin, hygromycin, kanamycin, mitomycin, mycophenolic acid, nalidixic acid, neomycin, nystatin, paromomycin, polymyxin, puromycin, rifampicin, spectinomycin, tetracycline, tylosin, and zeocin. Also contemplated is the supplementation of cell culture medium with mammalian plasma or sera. Plasma or sera often contain cellular factors and components that are necessary for viability and expansion. The use of suitable serum replacements is also contemplated.

[0095] Reference to particular buffers, media, reagents, cells, culture conditions, and the like, or to some subclass of same, is not intended to be limiting but should be read to include all such related materials that one of ordinary skill in the art would recognize as being of interest or value in the particular context in which that discussion is presented. For example, it is often possible to substitute one buffer system or culture medium for another, such that a different but known way is used to achieve the same goals as those to which the use of a suggested method, material, or composition is directed. In particular embodiments, cells are cultured in a cell culture system comprising a cell culture medium, preferably in a culture vessel, in particular, a cell culture medium supplemented with a substance suitable and determined for protecting the cells from in vitro aging and / or inducing in an unspecific or specific reprogramming.B. Cell Generation

[0096] Certain methods of the disclosure concern culturing the cells obtained from human tissue samples. In particular embodiments of the present disclosure, cells are plated onto a substrate that allows for adherence of cells thereto. This may be carried out, for example, by plating the cells in a culture plate that displays one or more substrate surfaces compatible with cell adhesion. When one or more substrate surfaces contact the suspension of cells (e.g., suspension in a medium) introduced into the culture system, cell adhesion between the cells and the substrate surfaces may ensue. Accordingly, in certain embodiments, cells are introduced into a culture system that features at least one substrate surface that is generally compatible with the adherence of cells thereto, such that the plated cells can contact the said substrate surface; such embodiments encompass plating onto a substrate, which allows adherence of cells thereto.

[0097] Cells of the present disclosure may be identified and characterized by their expression of specific marker proteins, such as cell surface markers. Detection and isolation of these cells can be achieved, for example, through flow cytometry, ELISA, and / or magnetic beads. Reverse-transcription polymerase chain reaction (RT-PCR) may be used to quantify cell-specific genes and / or to monitor changes in gene expression in response to differentiation. In certain embodiments, the marker proteins used to identify and characterize the cells are selected from the list consisting of c-Kit, Nanog, Sox2, Heyl, SMA, Vimentin, Cyclin D2, Snail, E-cadherin, Nkx2.5, GATA4, CD105, CD90, CD29, CD73, Wtl, CD34, CD45, and a combination thereof.C. Pharmaceutical Compositions

[0098] In certain aspects, the compositions or agents for use in the methods, such as fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast microvesicles, fibroblast lysate, fibroblast apoptotic bodies, fibroblast conditioned media, or a combination thereof, are suitably contained in a pharmaceutically acceptable carrier. The carrier is non-toxic and biocompatible and is selected so as not to detrimentally affect the biological activity of the agent. The agents in some aspects of the disclosure may be formulated into preparations for local delivery (z.e., to a specific location of the body, such as on or in the skin or other tissue) or systemic delivery, in solid, semi-solid, gel, liquid or gaseous forms such as tablets, capsules, powders, granules, ointments, solutions, depositories, inhalants and injections allowing for oral, parenteral or surgical administration. Certain aspects of the disclosure also contemplate local administration of the compositions by coating medical devices and the like.

[0099] Suitable carriers for parenteral delivery via injectable, infusion, or irrigation and topical delivery include distilled water, physiological phosphate-buffered saline, normal or lactated Ringer's solutions, dextrose solution, Hank's solution, or propanediol. In addition, sterile, fixed oils may be employed as a solvent or suspending medium. For this purpose, any biocompatible oil may be employed, including synthetic mono- or di-glycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables. The carrier and agent may be compounded as a liquid, suspension, polymerizable or non-polymerizable gel, paste, or salve.

[0100] The carrier may also comprise a delivery vehicle to sustain (i.e., extend, delay, or regulate) the delivery of the agent(s) or to enhance the delivery, uptake, stability, or pharmacokinetics of the therapeutic agent(s). Such a delivery vehicle may include, by way of non-limiting examples, microparticles, microspheres, nanospheres or nanoparticles composed of proteins, liposomes, carbohydrates, synthetic organic compounds, inorganic compounds, polymeric or copolymeric hydrogels, and polymeric micelles.

[0101] In certain aspects, the actual dosage amount of a composition administered to a patient or subject can be determined by physical and physiological factors such as body weight, severity of the condition, the type of disease being treated, previous or concurrent therapeutic interventions, idiopathy of the patient and the route of administration. The practitioner responsible for administration will, in any event, determine the concentration of active ingredient(s) in a composition and appropriate dose(s) for the individual subject.

[0102] Solutions of pharmaceutical compositions can be prepared in water suitably mixed with a surfactant, such as hydroxypropyl cellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, mixtures thereof, and oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0103] In certain aspects, the pharmaceutical compositions are advantageously administered in the form of injectable compositions either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid prior to injection may also be prepared. These preparations also may be emulsified. A typical composition for such purpose comprises a pharmaceutically acceptable carrier. For instance, the composition may contain 10 mg or less, 25 mg, 50 mg or up to about 100 mg of human serum albumin per milliliter of phosphate-buffered saline. Other pharmaceutically acceptable carriers include aqueous solutions and non-toxic excipients, including salts, preservatives, buffers, and the like.

[0104] Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, saline solutions, and parenteral vehicles such as sodium chloride, Ringer's dextrose, etc. Intravenous vehicles include fluid and nutrient replenishers. Preservatives include antimicrobial agents, antifungal agents, antioxidants, chelating agents, and inert gases. The pH and exact concentration of the various components of the pharmaceutical composition are adjusted according to well-known parameters.

[0105] Additional formulations are suitable for oral administration. Oral formulations include such typical excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and the like. The compositions take the form of solutions, suspensions, tablets, pills, capsules, sustained release formulations or powders.

[0106] In further aspects, the pharmaceutical compositions may include classic pharmaceutical preparations. Administration of pharmaceutical compositions according to certain aspects may be via any common route so long as the target tissue is available via that route. This may include oral, nasal, buccal, rectal, vaginal, or topical. Alternatively, administration may be by orthotopic, intradermal, subcutaneous, intramuscular, intraperitoneal, or intravenous injection. Such compositions would normally be administered as pharmaceutically acceptable compositions that include physiologically acceptable carriers, buffers, or other excipients. For the treatment of conditions of the lungs, aerosol delivery can be used. The volume of the aerosol may be between about 0.01 ml and 0.5 ml, for example.

[0107] An effective amount of the pharmaceutical composition is determined based on the intended goal. The term “unit dose” or “dosage” refers to physically discrete units suitable for use in a subject, each unit containing a predetermined quantity of the pharmaceutical composition calculated to produce the desired responses discussed above in association with its administration, i.e., the appropriate route and treatment regimen. The quantity to be administered, both according to the number of treatments and unit dose, depends on the protection or effect desired.

[0108] Precise amounts of the pharmaceutical composition also depend on the judgment of the practitioner and are peculiar to each individual. Factors affecting the dose include the physical and clinical state of the patient, the route of administration, the intended goal of treatment (e.g., alleviation of symptoms versus cure) and the potency, stability, and toxicity of the particular therapeutic substance.II. Formulations and Culture of the Cells

[0109] In particular embodiments, the cells of the disclosure may be specifically formulated, and / or they may be cultured in a particular medium. The cells may be formulated in such a manner as to be suitable for delivery to a recipient without deleterious effects.

[0110] The medium in certain aspects can be prepared using a medium used for culturing animal cells as their basal medium, such as any of AIM V, X-VIVO-15, NeuroBasal, EGM2, TeSR, BME, BGJb, CMRL 1066, Glasgow MEM, Improved MEM Zinc Option, IMDM, Medium 199, Eagle MEM, aMEM, DMEM, Ham, RPMI-1640, and Fischer's media, as well as any combinations thereof, but the medium may not be particularly limited thereto as far as it can be used for culturing animal cells. Particularly, the medium may be xeno-free or chemically defined.[OHl] The medium can be a serum-containing or serum-free medium, or xeno-free medium. From the aspect of preventing contamination with heterogeneous animal-derived components, serum can be derived from the same animal as that of the stem cell(s). The serum- free medium refers to a medium with no unprocessed or unpurified serum and, accordingly, can include a medium with purified blood-derived components or animal tissue-derived components (such as growth factors).

[0112] The medium may contain or may not contain any alternatives to serum. The alternatives to serum can include materials that appropriately contain albumin (such as lipid- rich albumin, bovine albumin, albumin substitutes such as recombinant albumin or a humanized albumin, plant starch, dextrans, and protein hydrolysates), transferrin (or other iron transporters), fatty acids, insulin, collagen precursors, trace elements, 2-mercaptoethanol, 3'- thiolgiycerol, or equivalents thereto. The alternatives to serum can be prepared using the method disclosed in International Publication No. 98 / 30679, for example (incorporated herein in its entirety). Alternatively, any commercially available materials can be used for more convenience. The commercially available materials include knockout Serum Replacement (KSR), Chemically-defined Lipid concentrated (Gibco), and Glutamax (Gibco).

[0113] In certain embodiments, the medium may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more of the following: Vitamins such as biotin; DL Alpha Tocopherol Acetate; DL Alpha-Tocopherol; Vitamin A (acetate); proteins such as BSA (bovine serum albumin) or human albumin, fatty acid-free Fraction V; Catalase; Human Recombinant Insulin; Human Transferrin; Superoxide Dismutase; Other Components such as Corticosterone; D-Galactose; Ethanolamine HC1; Glutathione (reduced); L-Carnitine HC1;Linoleic Acid; Linolenic Acid; Progesterone; Putrescine 2HC1; Sodium Selenite; and / or T3 (triodo-I-thyronine). In certain embodiments, one or more of these may be explicitly excluded.

[0114] In some embodiments, the medium further comprises vitamins. In some embodiments, the medium comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 of the following (and any range derivable therein): biotin, DL alpha-tocopherol acetate, DL alpha-tocopherol, vitamin A, choline chloride, calcium pantothenate, pantothenic acid, folic acid nicotinamide, pyridoxine, riboflavin, thiamine, inositol, vitamin B12, or the medium includes combinations thereof or salts thereof. In some embodiments, the medium comprises or consists essentially of biotin, DL alpha-tocopherol acetate, DL alpha-tocopherol, vitamin A, choline chloride, calcium pantothenate, pantothenic acid, folic acid nicotinamide, pyridoxine, riboflavin, thiamine, inositol, and vitamin B 12. In some embodiments, the vitamins include or consist essentially of biotin, DL alpha-tocopherol acetate, DL alpha-tocopherol, vitamin A, or combinations or salts thereof. In some embodiments, the medium further comprises proteins. In some embodiments, the proteins comprise albumin or bovine serum albumin, a fraction of BSA, catalase, insulin, transferrin, superoxide dismutase, or combinations thereof. In some embodiments, the medium further comprises one or more of the following: corticosterone, D-Galactose, ethanolamine, glutathione, L-camitine, linoleic acid, linolenic acid, progesterone, putrescine, sodium selenite, or triodo-I-thyronine, or combinations thereof. In some embodiments, the medium comprises one or more of the following: a B-27® supplement, xeno-free B-27® supplement, GS21™ supplement, or combinations thereof. In some embodiments, the medium comprises or further comprises amino acids, monosaccharides, inorganic ions. In some embodiments, the amino acids comprise arginine, cystine, isoleucine, leucine, lysine, methionine, glutamine, phenylalanine, threonine, tryptophan, histidine, tyrosine, or valine, or combinations thereof. In some embodiments, the inorganic ions comprise sodium, potassium, calcium, magnesium, nitrogen, or phosphorus, or combinations or salts thereof. In some embodiments, the medium further comprises one or more of the following: molybdenum, vanadium, iron, zinc, selenium, copper, or manganese, or combinations thereof. In certain embodiments, the medium comprises or consists essentially of one or more vitamins discussed herein and / or one or more proteins discussed herein, and / or one or more of the following: corticosterone, D-Galactose, ethanolamine, glutathione, L-carnitine, linoleic acid, linolenic acid, progesterone, putrescine, sodium selenite, or triodo-I-thyronine, a B-27® supplement, xeno-free B-27® supplement, GS21TM supplement, an amino acid (such as arginine, cystine, isoleucine, leucine, lysine, methionine, glutamine, phenylalanine, threonine, tryptophan, histidine, tyrosine, or valine), monosaccharide, inorganic ion (such as sodium, potassium, calcium, magnesium, nitrogen,and / or phosphorus) or salts thereof, and / or molybdenum, vanadium, iron, zinc, selenium, copper, or manganese. In certain embodiments, one or more of these may be explicitly excluded.

[0115] The medium can also contain one or more externally added fatty acids or lipids, amino acids (such as non-essential amino acids), vitamin(s), growth factors, cytokines, antioxidant substances, 2-mercaptoethanol, pyruvic acid, buffering agents, and / or inorganic salts. In certain embodiments, one or more of these may be explicitly excluded.

[0116] One or more of the medium components may be added at a concentration of at least, at most, or about 0.1, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 180, 200, 250 ng / L, ng / ml, pg / ml, mg / ml, or any range derivable therein.

[0117] In certain embodiments, the cells of the disclosure are certain ally formulated. They may or may not be formulated as a cell suspension. In certain cases, they are formulated in a single dose form. They may be formulated for systemic or local administration. In some cases, the cells are formulated for storage prior to use, and the cell formulation may comprise one or more cryopreservation agents, such as DMSO (for example, in 5% DMSO). The cell formulation may comprise albumin, including human albumin, with a specific formulation comprising 2.5% human albumin. The cells may be formulated specifically for intravenous administration; for example, they are formulated for intravenous administration over less than one hour. In particular embodiments, the cells are in a formulated cell suspension that is stable at room temperature for 1, 2, 3, or 4 hours or more from the time of thawing.IV. Kits of the Disclosure

[0118] Any of the cellular and / or non-cellular compositions described herein or similar thereto may be comprised in a kit. In a non-limiting example, one or more reagents for use in methods for preparing cellular therapy may be comprised in a kit. Such reagents may include cells, IFN-gamma, platelet-rich plasma, platelet lysate, one or more angiogenic factors, one or more growth factors, vector(s) one or more costimulatory factors, media, enzymes, buffers, nucleotides, salts, primers, and so forth. The kit may comprise any protein listed in the disclosure. The kit components are provided in suitable container means.

[0119] Some components of the kits may be packaged either in aqueous media or in lyophilized form. The container means of the kits will generally include at least one vial, test tube, flask, bottle, syringe, or other container means, into which a component may be placed, and preferably, suitably aliquoted. Where there is more than one component in the kit, the kitalso will generally contain a second, third, or other additional container into which the additional components may be separately placed. However, various combinations of components may be comprised in a vial. The kits of the present disclosure also will typically include a means for containing the components in close confinement for commercial sale. Such containers may include injection or blow-molded plastic containers into which the desired vials are retained.

[0120] When the components of the kit are provided in one and / or more liquid solutions, the liquid solution is an aqueous solution, with a sterile aqueous solution being particularly useful. In some cases, the container means may itself be a syringe, pipette, and / or other such like apparatus, or may be a substrate with multiple compartments for a desired reaction.

[0121] Some components of the kit may be provided as dried powder(s). When reagents and / or components are provided as dry powder, the powder can be reconstituted by the addition of a suitable solvent. It is envisioned that the solvent may also be provided in another container means. The kits may also comprise a second container means for containing a sterile acceptable buffer and / or other diluent.

[0122] In certain embodiments, reagents and materials include primers for amplifying desired sequences, nucleotides, suitable buffers or buffer reagents, salt, and so forth, and in some cases, the reagents include apparatus or reagents for isolation of a particular desired cell(s).

[0123] In particular embodiments, there are one or more apparatuses in the kit suitable for extracting one or more samples from an individual. The apparatus may be a syringe, fine needles, scalpel, and so forth.V. Examples

[0124] The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the certain embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the disclosure.EXAMPLE 1TREATMENT OF PSORIASIS

[0125] Animals and psoriasis disease model. For psoriasis induction on the back, mice were anesthetized using an isoflurane respirator. A rectangular area of 3x4-cm was shaved on the back skin, and the remaining hairs were removed using depilatory cream (Veet, USA). Psoriasis was induced via topical application of 31.25 mg of iMQ cream (5% IMQ, Taro Pharmaceutical, Hawthorne, NY, USA) to the lower dorsal skin for 8 or 15 consecutive days. Mouse body weight was recorded daily. All experimental procedures followed the NIH Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee at the K2Bio Lab (Houston, TX, USA).

[0126] Clinical assessment of psoriasis. Phenotypic features of IMQ-induced psoriasis were assessed daily using the Psoriasis Area and Severity Index (PASI). Briefly, double-folded skin thickness was assessed using a digital micrometer (Rexbeti, Auburn, Washington, USA). Dorsal skin parameters, including thickness, erythema, and scaling, were scored daily on a scale of 0 (no incidence of disease) to 4 (most severe), respectively. The three-parameter scores were then averaged to generate the daily average score for each mouse. Tail vein intravenous injection of fibroblast spheroids was performed when the average clinical score reached 2. Both groups continued to be treated with the IMQ cream in order to maintain the psoriasis-induction pressure on the skin. Mice were terminated when the average clinical score in the IMQ-treated control group reached 3.5.

[0127] Psoriasis Area and Severity Index (PASI):

[0128] Cell culture umi treatment. Fibroblast cells were used at passages 5-7 and maintained in fibroblast growth medium at 37°C with 5% CO2 until 70-80% confluence. For preparing Human Dermal Fibroblast (HDF) spheroids, HDF were trypsinized using 0.25% trypsin for 5 min. HDF spheroids were cultured using Elplasia® microcavity plates for 3-4 days. HDF spheroids (577 of 150 pm, each containing 1,733 cells, totaling IxlO6cells per mouse) were administered via tail vein either as a single dose at mild psoriasis (PASI score 2) or multiple doses (every 3 days) at moderate-to-severe psoriasis (PASI score 4). Blood analysis included complete blood count (CBC) and psoriatic-related cytokine levels using Luminex assays. Skin sections were analyzed for epidermal thickness, immune cell infiltration, and cell proliferation.

[0129] Statistical analysis. Statistical analysis was performed using GraphPad Prism version 9 (GraphPad Software, La Jolla, CA, USA). Results are expressed as the mean ± SEM. Normality tests were performed using D’Agostino-Pearson. The statistical difference between groups was analyzed using one-way ANOVA tests followed by post-hoc comparisons. A p value of less than 0.05 was considered statistically significant.

[0130] Results. Results are depicted in Figures 1 - 10 and summarized in the BRIEF DESCRIPTION OF THE DRAWINGS section.REFERENCES

[0131] The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference.1. Clark, R.A. and T.S. Kupper, IL-15 and dermal fibroblasts induce proliferation of natural regulatory T cells isolated from human skin. Blood, 2007. 109(1): p. 194-202.2. Bakar, R.S., et al., Depression and anxiety among patients with psoriasis: A correlation with quality of life and associated factors. J Taibah Univ Med Sci, 2021. 16(4): p. 491-496.3. Gisondi, P., et al., Pathogenesis of Chronic Plaque Psoriasis and Its Intersection With Cardio-Metabolic Comorbidities. Front Pharmacol, 2020. 11: p. 117.4. Zhang, H., et al., Stigmatization in Patients With Psoriasis: A Mini Review. Front Immunol, 2021. 12: p. 715839.5. Armstrong, A.W. and C. Read, Pathophysiology, Clinical Presentation, and Treatment of Psoriasis: A Review. Jama, 2020. 323(19): p. 1945-1960.6. Lowes, M.A., M. Suarez-Farinas, and J.G. Krueger, Immunology of psoriasis. Annu Rev Immunol, 2014. 32: p. 227-55.7. Rendon, A. and K. Schakel, Psoriasis Pathogenesis and Treatment. Int J Mol Sci, 2019. 20(6).8. Samotij, D., et al., Pathogenesis of psoriasis in the "omic" era. Part I. Epidemiology, clinical manifestation, immunological and neuroendocrine disturbances. Postepy Dermatol Alergol, 2020. 37(2): p. 135-153.9. Thangapazham, R.L., T.N. Darling, and J. Meyerle, Alteration of skin properties with autologous dermal fibroblasts. Int J Mol Sci, 2014. 15(5): p. 8407-27.10. Kim, S., et al., Genome editing of immune cells using CRISPR / Cas9. BMB Rep, 2021. 54(1): p. 59-69.11. Basu, A. and V.K. Tiwari, Epigenetic reprogramming of cell identity: lessons from development for regenerative medicine. Clin Epigenetics, 2021. 13(1): p. 144.* * *

[0132] All of the methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit, and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.ADDITIONAL RECITED EMBODIMENTS:

[0133] Embodiment 1 : A method of treating psoriasis in an individual, comprising the step of administering to the individual a therapeutically effective amount of a composition comprising fibroblasts and / or fibroblast-derived products.

[0134] Embodiment 2: The method of embodiment 1, wherein the fibroblast-derived products comprise fibroblast spheroids, fibroblast exosomes, fibroblast microvesicles, fibroblast lysate, fibroblast apoptotic bodies, fibroblast conditioned media, or any combination thereof.

[0135] Embodiment 3 : The method of embodiment 1 or 2, wherein the composition reduces the proliferation of psoriasis-associated immune cells.

[0136] Embodiment 4: The method of embodiment 4, wherein the psoriasis-associated immune cells express pathogenic levels of IL-23, IL-1, IL-4, IL-10, IL-14, IL-22, IL-17, IL-12. TNF, INF- y, TGF-pi, or a combination thereof.

[0137] Embodiment 5: The method of embodiment 3 or 4, wherein the psoriasis-associated immune cells are pathogenic T cells, pathogenic dendritic cells, pathogenic macrophages, pathogenic neutrophils, or any combination thereof.

[0138] Embodiment 6: The method of any one of embodiments 1-3, wherein the composition is administered topically, intravenously, intrathecally, or by subcutaneous injection.

[0139] Embodiment 7: The method of any one of embodiments 1-6, wherein the composition comprises fibroblasts or fibroblast-derived products that are autologous with respect to the individual.

[0140] Embodiment 8: The method of any one of embodiments 1-6, wherein the composition comprises fibroblasts or fibroblast-derived products that are allogeneic with respect to the individual.

[0141] Embodiment 9: The method of any one of embodiments 1-6, wherein the composition comprises fibroblasts or fibroblast-derived products that are xenogeneic with respect to the individual.

[0142] Embodiment 10: The method of any one of embodiments 1-6, wherein the composition comprises fibroblasts or fibroblast-derived products that are syngeneic with respect to the individual.

[0143] Embodiment 11 : A method of treating psoriasis in an individual, comprising the step of providing to the individual a composition with an effective amount of: (1) fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials; and optionally (2) one or more types of immune cells and / or one or more derivative agents of the immune cells, optionally wherein the derivative agents are cytokines and / or chemokines and / or growth factors.

[0144] Embodiment 12: The method of embodiment 11, wherein the providing the composition comprises placing the composition (a) into the blood stream of the individual and / or (b) into and / or onto the skin tissue of the individual, optionally via intravenous injection or subcutaneous injection.

[0145] Embodiment 13: The method of embodiment 11 or 12, wherein the composition decreases the proliferation of one or more types of pathogenic immune cells.

[0146] Embodiment 14: The method of any one of embodiments 11-13, wherein the providing of (1) and (2) occurs generally simultaneously.

[0147] Embodiment 15: The method of any one of embodiments 11-13, wherein the providing of (1) and (2) occurs at different times.

[0148] Embodiment 16: The method of embodiment 15, wherein (1) occurs before (2).

[0149] Embodiment 17: The method of embodiment 15, wherein (1) occurs after (2).

[0150] Embodiment 18: The method of embodiment 11, wherein the composition comprises activated fibroblasts and / or the fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, or fibroblast-derived materials come from activated fibroblasts.

[0151] Embodiment 19: The method of embodiment 12, wherein the composition comprises fibroblasts that are activated ex vivo with one or more nucleic acids, one or more transcription factor activators, one or more growth factors, one or more cytokines, one or more chemokines, and / or with co-culture with non-fibroblast cells, optionally wherein the fibroblasts are activated ex vivo with IL-1, IL-6, IL-8, MCP-1, CCL-2, prostaglandins, miRNA-21, miRNA-23b, miRNA-31, miR-215, miR-1, miR-133, miR-208, miR-499, transcription factors Mcm3, Dicerl, Cdc25A, Ick, Tripl3, McmlO, OCT4, SOX2, KLF4, MYC, Pax3, or a combination thereof.

[0152] Embodiment 20: The method of embodiment 19, wherein the non-fibroblast cells are one or more types of immune cells.

[0153] Embodiment 21 : The method of embodiment 20, wherein the immune cells are T cells, dendritic cells, natural killer cells, natural killer T cells, macrophages, microglia, B cells, neutrophils, or a combination thereof.

[0154] Embodiment 22: A method of treating psoriasis in an individual, comprising the step of providing to the individual a composition with an effective amount of: (1) fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials; wherein the fibroblasts are activated with one or more agents; and / or the fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, or fibroblast-derived materials come from activated fibroblasts activated with one or more agents; and (2) one or more types of immune cells and / or one or more derivative agents thereof, optionally wherein the derivative agents are cytokines and / or chemokines and / or growth factors.

[0155] Embodiment 23: The method of embodiment 22, wherein the composition comprising fibroblasts is activated ex vivo with one or more nucleic acids, one or more transcription factor activators, one or more growth factors, one or more cytokines, one or more chemokines, and / or with co-culture with non-fibroblast cells.

[0156] Embodiment 24: The method of embodiment 22, wherein the providing of the composition comprises administering the composition (a) into the blood stream of the individual and / or (b) into and / or onto the skin tissue of the individual.

[0157] Embodiment 25: The method of embodiment 22 or 23, wherein following providing the composition to the individual, there is a decrease of the expression in immune cells of the individual of pathogenic levels from the cells of one or more of IL-23, IL-1, IL-10, IL-22, IL- 17, IL- 12, TNF, INF-y, and / or TGF-pi.

[0158] Embodiment 26: A method of epigenetic reprogramming of a composition comprising localized fibroblasts, keratinocytes, and / or epithelial cells in an individual with psoriasis, comprising the step of providing the individual an effective amount of: (1) fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials; wherein the fibroblasts are activated with one or more agents; and / or the fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, or fibroblast-derived materials come from activated fibroblasts activated with one or more agents; and (2) one or more types of immune cells or derivative agents.

[0159] Embodiment 27: The method of embodiment 26, wherein the providing of the composition comprises placing the composition (a) into the blood stream of the individual and / or (b) into and / or onto and / or near the skin tissue of the individual.

[0160] Embodiment 28: A method of enabling the activation and migration of localized stem cell niches to replace the pathogenetic keratinocytes, fibroblasts, and / or epithelial cells in the skin of an individual, comprising the step of providing to the individual a composition comprising an effective amount of: (1) fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials; wherein the fibroblasts are activated with one or more agents and / or the fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, or fibroblast-derived materials come from activated fibroblasts activated with one or more agents; and / or (2) one or more types of immune cells or derivative agents.

[0161] Embodiment 29: The method of embodiment 28, wherein the providing the composition comprises placing the composition (a) into the blood stream of the individual and / or (b) into and / or onto and / or near the skin tissue of the individual.

[0162] Embodiment 30: A method of eliciting an immune response in an individual with psoriasis, comprising the step of providing to the individual a composition comprising an effective amount of: (1) fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials; and (2) one or more adjuvants.

[0163] Embodiment 31 : The method of embodiment 30, wherein the adjuvant is chemical product-based, viral product-based, bacterial product-based, or a mixture thereof.

[0164] Embodiment 32: The method of any one of embodiments 1-31, further comprising administering one or more adjuvants to the individual, wherein the adjuvant comprises one or more of steroid cream, roflumilast, moisturizer, Salicylic acid, Coal tar, Vitamin D-based cream, Vitamin D-based ointment, Retinoid cream, Calcineurin inhibitor, antibiotic, Anthralin, and / or any combination thereof.

[0165] Embodiment 33: The method of any one of embodiments 1-32, wherein the composition is administered intravenously, topically, intramuscularly, subcutaneously, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, intranasally or any combination thereof.

[0166] Embodiment 34: The method of any one of embodiments 1-33, wherein the composition is administered to the individual intravenously, topically, intramuscularly, subcutaneously, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally.

[0167] Embodiment 35: The method of any one of embodiments 1-34, wherein the composition comprises liquid solutions or suspensions, solid forms suitable for use to prepare solutions, and / or suspensions upon the addition of a liquid prior to injection.

[0168] Embodiment 36: The method of any one of embodiments 1-35, wherein the composition is in an injectable form and comprises a sterile aqueous solution or dispersion comprising aqueous propylene glycol or comprises a sterile powder for the extemporaneous preparation of sterile injectable solution or dispersion.

[0169] Embodiment 37: The method of any one of embodiments 1-36, wherein the composition comprises a neutral or salt form comprising salts, wherein the salts optionally comprise acid addition salts comprising free amino groups of one or more proteins; optionally wherein the acid addition salts are formed with one or more inorganic acids and / or one or more organic acids.

[0170] Embodiment 38: The method of any one of embodiments 1-37, wherein the composition further comprises a salt formed with one or more free carboxyl groups, wherein the one or more free carboxyl groups are derived from (a) inorganic bases comprising one or more of sodium, potassium, ammonium, calcium, or ferric hydroxides; and / or (b) organic bases comprising one or more of isopropylamine, trimethylamine, histidine, procaine, and / or any combination thereof.

[0171] Embodiment 39: The method of any one of embodiments 1-38, wherein the composition comprises a solvent and dispersion medium comprising one or more of water,vegetable oils, ethanol, polyol comprising glycerol, propylene glycol, liquid polyethylene glycol, and / or any combination thereof.

[0172] Embodiment 40: The method of any one of embodiments 1-39, wherein the composition comprising fibroblast cells comprises: fibroblast cells cultured for at least between about 10 days and about 40 days, optionally for at least between about 15 days and about 35 days, wherein the fibroblasts are cultured in the presence of a liquid culture medium, wherein the liquid culture medium comprises a basal medium formulation comprising one or more of Eagle's Minimum Essential Medium (MEM), Dulbecco's Modified Eagle's Medium (DMEM), alpha modified Minimum Essential Medium (alpha-MEM), Basal Medium Essential (BME), Iscove's Modified Dulbecco's Medium (IMDM), BGJb medium, F-12 Nutrient Mixture (Ham), Liebovitz L-15, DMEM / F-12, Essential Modified Eagle's Medium (EMEM), RPMI-1640, and / or modifications, and / or any combination thereof.

[0173] Embodiment 41 : The method of embodiment 40, wherein the liquid culture medium comprises Dulbecco's Modified Eagle's Medium (DMEM), alpha-modified Minimal Essential Medium (aMEM), and Roswell Park Memorial Institute Media 1640 (RPMI Media 1640), and / or any combination thereof.

[0174] Embodiment 42: The method of any one of embodiment 40 or 41, wherein the liquid culture medium further comprises 20% fetal bovine serum (FBS) or 1-20% horse serum above a standard liquid culture medium.

[0175] Embodiment 43 : The method of any one of embodiments 40-42, wherein the liquid culture medium further comprises an additional supplement to supply the cells with one or more of antioxidant supplement, necessary trace element, and / or substance for optimal growth and / or expansion; optionally wherein the additional supplement comprises one or more of insulin, transferrin, selenium salts, and / or any combination thereof; optionally wherein the additional supplements are included in one or more of a salt solution Hanks' Balanced Salt Solution (HBSS), and / or Earle's Salt Solution; optionally wherein antioxidant supplement comprises P-mercaptoethanol.

[0176] Embodiment 44: The method of any one of embodiments 40-43, wherein the liquid culture medium further comprises an antibiotic and / or antimycotic compound; optionally wherein the antibiotic and / or antimycotic compound comprises penicillin, streptomycin, and / or another compound(s); optionally wherein the antibiotic and / or antimycotic compound comprises amphotericin, ampicillin, gentamicin, bleomycin, hygromycin, kanamycin, mitomycin, mycophenolic acid, nalidixic acid, neomycin, nystatin, paromomycin, polymyxin,puromycin, rifampicin, spectinomycin, tetracycline, tylosin, zeocin, and / or any combination thereof.

[0177] Embodiment 45: The method of any one of embodiments 1-44, wherein the cells comprising the composition are grown: at temperatures between 27° C to 40° C, such as 31° C to 37° C; with a humidified incubator; with a carbon dioxide content maintained between 2% to 10%; and / or with an oxygen content maintained between 1% and 22%.

[0178] Embodiment 46: The method of any one of embodiments 1-45, wherein the composition is suitably contained in a pharmaceutically acceptable carrier delivered to the individual by methods comprising one or more of injectable, infusion, irrigation, and / or topical delivery; and optionally wherein the pharmaceutically acceptable carrier comprises one or more of distilled water, physiological phosphate-buffered saline, normal or lactated Ringer's solutions, dextrose solution, Hank's solution, propanediol, sterile and / or fixed oils employed as a solvent or suspending medium, and / or any combination thereof.

[0179] Embodiment 47: The method of embodiment 46, wherein the pharmaceutically acceptable carrier comprises a delivery vehicle to extend, delay, or regulate the delivery of the composition or to enhance the delivery, uptake, stability, or pharmacokinetics of the therapeutic agents within the composition.

[0180] Embodiment 48: The method of any one of embodiments 1-47, wherein the pharmaceutically acceptable carrier comprises microparticles, microspheres, nanospheres, and / or nanoparticles; optionally, wherein the pharmaceutically acceptable carrier comprises one or more of proteins, liposomes, carbohydrates, synthetic organic compounds, inorganic compounds, polymeric or copolymeric hydrogels, and / or polymeric micelles, and / or any combination thereof.

[0181] Embodiment 49: The method of any one of embodiments 1-48, wherein the composition is in an injectable form, wherein the injectable form is emulsified, optionally wherein the injectable form comprises 0.1 mg or up to about 100 mg of human serum albumin per milliliter of phosphate buffered saline.

[0182] Embodiment 50: The method of embodiment 49, wherein the composition in an injectable form comprises one or more of aqueous solutions, non-toxic excipients, salts, preservatives, buffers, and / or any combination thereof.

[0183] Embodiment 51 : The method of any one of embodiments 1-50, wherein the composition is an oral form, wherein the oral form comprises one or more of pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and / or any combination thereof and optionally wherein the oral form composition takes the form of one or more of a solution, suspension, tablet, pill, capsule, sustained release formulation, powder and / or any combination thereof.

[0184] Embodiment 52: The method of any one of embodiments 1-51, wherein the cells of the composition are cultured in a medium, wherein the medium is prepared using a medium used for culturing animal cells as their basal medium, optionally wherein the medium comprises one or more of AIM V, X-VIVO-15, NeuroBasal, EGM2, TeSR, BME, BGJb, CMRL 1066, Glasgow MEM, Improved MEM Zinc Option, IMDM, Medium 199, Eagle MEM, aMEM, DMEM, Ham, RPMI-1640, and / or Fischer's media, and / or any combination thereof.

[0185] Embodiment 53: The method of embodiment 52, wherein the medium comprises serum-containing, serum-free, and / or xeno-free medium, or any combination thereof.

[0186] Embodiment 54: The method of any one of embodiments 1-53, wherein the composition is available in a kit comprising one or more reagents for use in methods for preparing cellular therapy.

[0187] Embodiment 55: The method of embodiment 54, wherein packaging of one or more reagents of the kit comprises aqueous media and the composition in lyophilized form.

[0188] Embodiment 56: The method of any one of embodiment 54 or 55, wherein the packaging of the one or more components of the kit further comprises liquid solutions and / or dried powders.

[0189] Embodiment 57: The method of any one of embodiments 1-56, wherein the fibroblasts are genetically engineered to attenuate their immune modulation potential, by modifying one or more genes.

[0190] Embodiment 58: The method of embodiment 57, wherein the one or more genes are selected from one or more of a gene for a cytokine, a soluble receptor, a mediator, and / or a cell surface receptor.

[0191] Embodiment 59: The method of embodiment 58, wherein the one or more genes are selected from: IL1RA, IL-4, IL-6, IL-10, IL-11, IL-13, TGFp, soluble TNF receptor p55, soluble TNF receptor p75, soluble IL-1 receptor type 2, membrane-bound IL-1 receptor type 2, IL- 18 binding protein, prostaglandin E2, PD1, CTLA4, TIM3, LAG3, TIGIT, CD96, BTLA, VISTA, Arginase, Prostaglandin E2, Indoleamin 2,3-dioxygenase, and / or Nitric oxide.

Claims

WHAT IS CLAIMED IS:

1. A method of treating psoriasis in an individual, comprising the step of administering to the individual a therapeutically effective amount of a composition comprising fibroblasts and / or fibroblast-derived products.

2. The method of claim 1, wherein the fibroblast-derived products comprise fibroblast spheroids, fibroblast exosomes, fibroblast microvesicles, fibroblast lysate, fibroblast apoptotic bodies, fibroblast conditioned media, or any combination thereof.

3. The method of claim 1 or 2, wherein the composition reduces the proliferation of psoriasis-associated immune cells.

4. The method of claim 4, wherein the psoriasis-associated immune cells express pathogenic levels of IL-23, IL-1, IL-4, IL-10, IL-14, IL-22, IL-17, IL-12, TNF, INF- y, TGF- pi, or a combination thereof.

5. The method of claim 3 or 4, wherein the psoriasis-associated immune cells are pathogenic T cells, pathogenic dendritic cells, pathogenic macrophages, pathogenic neutrophils, or any combination thereof.

6. The method of any one of claims 1-3, wherein the composition is administered topically, intravenously, intrathecally, or by subcutaneous injection.

7. The method of any one of claims 1-6, wherein the composition comprises fibroblasts or fibroblast-derived products that are autologous with respect to the individual.

8. The method of any one of claims 1-6, wherein the composition comprises fibroblasts or fibroblast-derived products that are allogeneic with respect to the individual.

9. The method of any one of claims 1-6, wherein the composition comprises fibroblasts or fibroblast-derived products that are xenogeneic with respect to the individual.

10. The method of any one of claims 1-6, wherein the composition comprises fibroblasts or fibroblast-derived products that are syngeneic with respect to the individual.

11. A method of treating psoriasis in an individual, comprising the step of providing to the individual a composition with an effective amount of:(1) fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials; and optionally(2) one or more types of immune cells and / or one or more derivative agents of the immune cells, optionally wherein the derivative agents are cytokines and / or chemokines and / or growth factors.

12. The method of claim 11, wherein the providing the composition comprises placing the composition (a) into the blood stream of the individual and / or (b) into and / or onto the skin tissue of the individual, optionally via intravenous injection or subcutaneous injection.

13. The method of claim 11 or 12, wherein the composition decreases the proliferation of one or more types of pathogenic immune cells.

14. The method of any one of claims 11-13, wherein the providing of (1) and (2) occurs generally simultaneously.

15. The method of any one of claims 11-13, wherein the providing of (1) and (2) occurs at different times.

16. The method of claim 15, wherein (1) occurs before (2).

17. The method of claim 15, wherein (1) occurs after (2).

18. The method of claim 11, wherein the composition comprises activated fibroblasts and / or the fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, or fibroblast-derived materials come from activated fibroblasts.

19. The method of claim 12, wherein the composition comprises fibroblasts that are activated ex vivo with one or more nucleic acids, one or more transcription factor activators, one or more growth factors, one or more cytokines, one or more chemokines, and / or with coculture with non-fibroblast cells, optionally wherein the fibroblasts are activated ex vivo with IL-1, IL-6, IL-8, MCP-1, CCL-2, prostaglandins, miRNA-21, miRNA-23b, miRNA-31, miR- 215, miR-1, miR-133, miR-208, miR-499, transcription factors Mcm3, Dicerl, Cdc25A, Ick, Tripl3, McmlO, OCT4, SOX2, KLF4, MYC, Pax3, or a combination thereof.

20. The method of claim 19, wherein the non-fibroblast cells are one or more types of immune cells.

21. The method of claim 20, wherein the immune cells are T cells, dendritic cells, natural killer cells, natural killer T cells, macrophages, microglia, B cells, neutrophils, or a combination thereof.

22. A method of treating psoriasis in an individual, comprising the step of providing to the individual a composition with an effective amount of:(1) fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials; wherein the fibroblasts are activated with one or more agents; and / or the fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, or fibroblast-derived materials come from activated fibroblasts activated with one or more agents; and(2) one or more types of immune cells and / or one or more derivative agents thereof, optionally wherein the derivative agents are cytokines and / or chemokines and / or growth factors.

23. The method of claim 22, wherein the composition comprising fibroblasts is activated ex vivo with one or more nucleic acids, one or more transcription factor activators, one or more growth factors, one or more cytokines, one or more chemokines, and / or with co-culture with non-fibroblast cells.

24. The method of claim 22, wherein the providing of the composition comprises administering the composition (a) into the blood stream of the individual and / or (b) into and / or onto the skin tissue of the individual.

25. The method of claim 22 or 23, wherein following providing the composition to the individual, there is a decrease of the expression in immune cells of the individual of pathogenic levels from the cells of one or more of IL-23, IL-1, IL-10, IL-22, IL-17, IL-12, TNF, INF-y, and / or TGF-pi.

26. A method of epigenetic reprogramming of a composition comprising localized fibroblasts, keratinocytes, and / or epithelial cells in an individual with psoriasis, comprising the step of providing the individual an effective amount of:(1) fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials; wherein the fibroblasts areactivated with one or more agents; and / or the fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, or fibroblast-derived materials come from activated fibroblasts activated with one or more agents; and(2) one or more types of immune cells or derivative agents.

27. The method of claim 26, wherein the providing of the composition comprises placing the composition (a) into the blood stream of the individual and / or (b) into and / or onto and / or near the skin tissue of the individual.

28. A method of enabling the activation and migration of localized stem cell niches to replace the pathogenetic keratinocytes, fibroblasts, and / or epithelial cells in the skin of an individual, comprising the step of providing to the individual a composition comprising an effective amount of(1) fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials; wherein the fibroblasts are activated with one or more agents and / or the fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, or fibroblast-derived materials come from activated fibroblasts activated with one or more agents; and / or(2) one or more types of immune cells or derivative agents.

29. The method of claim 28, wherein the providing the composition comprises placing the composition (a) into the blood stream of the individual and / or (b) into and / or onto and / or near the skin tissue of the individual.

30. A method of eliciting an immune response in an individual with psoriasis, comprising the step of providing to the individual a composition comprising an effective amount of(1) fibroblasts, fibroblast spheroids, fibroblast exosomes, fibroblast lysates, fibroblast apoptotic bodies, and / or fibroblast-derived materials; and(2) one or more adjuvants.

31. The method of claim 30, wherein the adjuvant is chemical product-based, viral productbased, bacterial product-based, or a mixture thereof.

32. The method of any one of claims 1-31, further comprising administering one or more adjuvants to the individual, wherein the adjuvant comprises one or more of steroid cream, roflumilast, moisturizer, Salicylic acid, Coal tar, Vitamin D-based cream, Vitamin D-based ointment, Retinoid cream, Calcineurin inhibitor, antibiotic, Anthralin, and / or any combination thereof.

33. The method of any one of claims 1-32, wherein the composition is administered intravenously, topically, intramuscularly, subcutaneously, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, intranasally or any combination thereof.

34. The method of any one of claims 1-33, wherein the composition is administered to the individual intravenously, topically, intramuscularly, subcutaneously, orally, transdermally, intraperitoneally, intraorbitally, by implantation, by inhalation, intrathecally, intraventricularly, or intranasally.

35. The method of any one of claims 1-34, wherein the composition comprises liquid solutions or suspensions, solid forms suitable for use to prepare solutions, and / or suspensions upon the addition of a liquid prior to injection.

36. The method of any one of claims 1-35, wherein the composition is in an injectable form and comprises a sterile aqueous solution or dispersion comprising aqueous propylene glycol or comprises a sterile powder for the extemporaneous preparation of sterile injectable solution or dispersion.

37. The method of any one of claims 1-36, wherein the composition comprises a neutral or salt form comprising salts, wherein the salts optionally comprise acid addition salts comprising free amino groups of one or more proteins; optionally wherein the acid addition salts are formed with one or more inorganic acids and / or one or more organic acids.

38. The method of any one of claims 1-37, wherein the composition further comprises a salt formed with one or more free carboxyl groups, wherein the one or more free carboxyl groups are derived from (a) inorganic bases comprising one or more of sodium, potassium, ammonium, calcium, or ferric hydroxides; and / or (b) organic bases comprising one or more of isopropylamine, trimethylamine, histidine, procaine, and / or any combination thereof.

39. The method of any one of claims 1-38, wherein the composition comprises a solvent and dispersion medium comprising one or more of water, vegetable oils, ethanol, polyol comprising glycerol, propylene glycol, liquid polyethylene glycol, and / or any combination thereof.

40. The method of any one of claims 1-39, wherein the composition comprising fibroblast cells comprises: fibroblast cells cultured for at least between about 10 days and about 40 days, optionally for at least between about 15 days and about 35 days, wherein the fibroblasts are cultured in the presence of a liquid culture medium, wherein the liquid culture medium comprises a basal medium formulation comprising one or more of Eagle's Minimum Essential Medium (MEM), Dulbecco's Modified Eagle's Medium (DMEM), alpha modified Minimum Essential Medium (alpha-MEM), Basal Medium Essential (BME), Iscove's Modified Dulbecco's Medium (IMDM), BGJb medium, F-12 Nutrient Mixture (Ham), Liebovitz L-15, DMEM / F-12, Essential Modified Eagle's Medium (EMEM), RPMI-1640, and / or modifications, and / or any combination thereof.

41. The method of claim 40, wherein the liquid culture medium comprises Dulbecco's Modified Eagle's Medium (DMEM), alpha-modified Minimal Essential Medium (aMEM), and Roswell Park Memorial Institute Media 1640 (RPMI Media 1640), and / or any combination thereof.

42. The method of any one of claim 40 or 41, wherein the liquid culture medium further comprises 20% fetal bovine serum (FBS) or 1-20% horse serum above a standard liquid culture medium.

43. The method of any one of claims 40-42, wherein the liquid culture medium further comprises an additional supplement to supply the cells with one or more of antioxidant supplement, necessary trace element, and / or substance for optimal growth and / or expansion; optionally wherein the additional supplement comprises one or more of insulin, transferrin, selenium salts, and / or any combination thereof; optionally wherein the additional supplements are included in one or more of a salt solution Hanks' Balanced Salt Solution (HBSS), and / or Earle's Salt Solution; optionally wherein antioxidant supplement comprises P-mercaptoethanol.

44. The method of any one of claims 40-43, wherein the liquid culture medium further comprises an antibiotic and / or antimycotic compound; optionally wherein the antibiotic and / or antimycotic compound comprises penicillin, streptomycin, and / or another compound(s); optionally wherein the antibiotic and / or antimycotic compound comprises amphotericin, ampicillin, gentamicin, bleomycin, hygromycin, kanamycin, mitomycin, mycophenolic acid, nalidixic acid, neomycin, nystatin, paromomycin, polymyxin, puromycin, rifampicin, spectinomycin, tetracycline, tylosin, zeocin, and / or any combination thereof.

45. The method of any one of claims 1-44, wherein the cells comprising the composition are grown: at temperatures between 27° C to 40° C, such as 31° C to 37° C; with a humidified incubator; with a carbon dioxide content maintained between 2% to 10%; and / or with an oxygen content maintained between 1% and 22%.

46. The method of any one of claims 1-45, wherein the composition is suitably contained in a pharmaceutically acceptable carrier delivered to the individual by methods comprising one or more of injectable, infusion, irrigation, and / or topical delivery; and optionally wherein the pharmaceutically acceptable carrier comprises one or more of distilled water, physiological phosphate-buffered saline, normal or lactated Ringer's solutions, dextrose solution, Hank's solution, propanediol, sterile and / or fixed oils employed as a solvent or suspending medium, and / or any combination thereof.

47. The method of claim 46, wherein the pharmaceutically acceptable carrier comprises a delivery vehicle to extend, delay, or regulate the delivery of the composition or to enhance the delivery, uptake, stability, or pharmacokinetics of the therapeutic agents within the composition.

48. The method of any one of claims 1-47, wherein the pharmaceutically acceptable carrier comprises microparticles, microspheres, nanospheres, and / or nanoparticles; optionally, wherein the pharmaceutically acceptable carrier comprises one or more of proteins, liposomes, carbohydrates, synthetic organic compounds, inorganic compounds, polymeric or copolymeric hydrogels, and / or polymeric micelles, and / or any combination thereof.

49. The method of any one of claims 1-48, wherein the composition is in an injectable form, wherein the injectable form is emulsified, optionally wherein the injectable form comprises 0.1 mg or up to about 100 mg of human serum albumin per milliliter of phosphate buffered saline.

50. The method of claim 49, wherein the composition in an injectable form comprises one or more of aqueous solutions, non-toxic excipients, salts, preservatives, buffers, and / or any combination thereof.

51. The method of any one of claims 1-50, wherein the composition is an oral form, wherein the oral form comprises one or more of pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and / or any combination thereof and optionally wherein the oral form composition takes the form of one or more of a solution, suspension, tablet, pill, capsule, sustained release formulation, powder and / or any combination thereof.

52. The method of any one of claims 1-51, wherein the cells of the composition are cultured in a medium, wherein the medium is prepared using a medium used for culturing animal cells as their basal medium, optionally wherein the medium comprises one or more of AIM V, X- VIVO-15, NeuroBasal, EGM2, TeSR, BME, BGJb, CMRL 1066, Glasgow MEM, Improved MEM Zinc Option, IMDM, Medium 199, Eagle MEM, aMEM, DMEM, Ham, RPMI-1640, and / or Fischer's media, and / or any combination thereof.

53. The method of claim 52, wherein the medium comprises serum-containing, serum-free, and / or xeno-free medium, or any combination thereof.

54. The method of any one of claims 1-53, wherein the composition is available in a kit comprising one or more reagents for use in methods for preparing cellular therapy.

55. The method of claim 54, wherein packaging of one or more reagents of the kit comprises aqueous media and the composition in lyophilized form.

56. The method of any one of claim 54 or 55, wherein the packaging of the one or more components of the kit further comprises liquid solutions and / or dried powders.

57. The method of any one of claims 1-56, wherein the fibroblasts are genetically engineered to attenuate their immune modulation potential, by modifying one or more genes.

58. The method of claim 57, wherein the one or more genes are selected from one or more of a gene for a cytokine, a soluble receptor, a mediator, and / or a cell surface receptor.

59. The method of claim 58, wherein the one or more genes are selected from: IL1RA, IL- 4, IL-6, IL-10, IL-11, IL-13, TGFP, soluble TNF receptor p55, soluble TNF receptor p75, soluble IL-1 receptor type 2, membrane-bound IL-1 receptor type 2, IL- 18 binding protein, prostaglandin E2, PD1, CTLA4, TIM3, LAG3, TIGIT, CD96, BTLA, VISTA, Arginase, Prostaglandin E2, Indoleamin 2, 3 -dioxygenase, and / or Nitric oxide.

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