KR101865040B1 - Use of mesenchymal stem cell having enhanced capability of producing immunoregulatory factor - Google Patents

Abstract

The present invention relates to a cytokine-treated mesenchymal stem cell, a method for enhancing inflammation regulatory factors of mesenchymal stem cell by cytokine treatment, and a composition for preventing and treating immunological diseases and inflammatory diseases comprising a mesenchymal stem cell having enhanced inflammation regulatory factors as an active ingredient. When culturing the mesenchymal stem cell, three or four cytokines are treated at low concentration to provide the mesenchymal stem cell enhanced in formation potential of inflammation regulatory factors, thereby maximizing the production of inflammation regulatory factors such as COX-2 and IDO to have a therapeutic effect on immunological diseases and inflammatory diseases.

Description

Use of mesenchymal stem cells having enhanced ability to produce immunomodulatory factors.

The present invention relates to the use of cytokines for enhancing the ability of mesenchymal stem cells to produce an inflammatory regulator and for the treatment of immune diseases and inflammatory diseases of mesenchymal stem cells enhanced in the ability to regulate inflammatory factors.

Stem cells are undifferentiated cells that can divide for a long time through self-renewal and can differentiate into various kinds of cells under specific circumstances. Stem cells can be divided into embryonic stem cells and adult stem cells depending on the origin of the stem cells, and there is a difficulty in ethical aspects and possibility of tumor formation by the treatment experiment using embryonic stem cells On the other hand, since adult stem cells can be easily obtained from various tissues, studies are being actively carried out to apply them to various diseases.

In particular, the most prominent feature of mesenchymal stem cells (MSC) is the ability to control inflammation, which is caused by soluble factors or direct contact between cells and cells. Interestingly, mesenchymal stem cells (MSCs) Is activated by an inflammatory cytokine such as interferon-gamma, IFN-gamma, tumor necrosis factor (TNF-a), etc., to induce anti-inflammatory factors, such as prostaglandin E2 , PGE2), interleukin-10, IL-10, HLA-G, nitiric oxide, indoleamine 2,3-dioxygenase And regulates the immune system by inducing the production of regulatory T cells (Treg) or inducing the death of immune cells involved in apoptosis.

 According to the report of the non-patent document 1, if the immune regulation of the mouse is mainly caused by nitric oxide, it has been proved that the case of human is manifested by IDO. In addition, PGE2 production is known to induce Foxp3 + Treg and type II macrophage cells, and to inhibit the function of NK cells and the maturation of dendritic cells.

 Accordingly, the present invention aims at maximizing the production of IDO and PGE2 in mesenchymal stem cells by low-level cytokine stimulation and contributing to development of immunoregulatory cells through mesenchymal stem cells.

Ren G et al. Stem Cells, Species Variation in the Mechanisms of Mesenchymal Stem Cell-Mediated Immunosuppression, Vol. 27, Issue 8, pp. 1954-1962, 2009

It is an object of the present invention to provide a use of three cytokines for enhancing the inflammatory regulator-producing ability of mesenchymal stem cells and for preventing or treating immunological diseases and inflammatory diseases of mesenchymal stem cells enhanced in productivity.

The inventors of the present invention have investigated the ability of the mesenchymal stem cell cytokine to produce an inflammatory regulator in combination with the cytokine of the mesenchymal stem cells. When the mesenchymal stem cells are cultured by treating three or four cytokines at a low concentration , Confirming that the cytokine enhances the ability of mesenchymal stem cells to regulate inflammatory factors, thereby completing the present invention.

Accordingly, the present invention provides three cytokines,

IFN-?;

TNF-a; And

IFN- ?, and IL-10. The present invention also relates to a composition for enhancing the inflammatory-modulating-factor-producing ability of mesenchymal stem cells.

The present invention also relates to a method for enhancing the ability of mesenchymal stem cells to produce an inflammatory regulator comprising the step of treating mesenchymal stem cells with a combination of three or four cytokines.

The present invention also relates to a pharmaceutical composition for the prevention or treatment of immunological diseases and inflammatory diseases, which comprises as an active ingredient mesenchymal stem cells enhanced in the ability to regulate inflammatory factors.

Hereinafter, the configuration of the present invention will be described in detail.

The present invention relates to a cytokine,

IFN-?;

TNF-a; And

A composition for enhancing the inflammatory-modulating-factor-producing ability of mesenchymal stem cells comprising at least one selected from IFN-β and IL-10,

Wherein each cytokine is contained in an amount of 1 to 50 ng / ml based on the number of mesenchymal stem cells 1 x ^{10 4} to 1 x 10 ⁶ cells.

In the present invention, " mesenchymal stem cells (MSCs) " are used in the same sense as adult stem cells, and include cord, cord blood, bone marrow, fat, muscle, nerve, And placenta. Among them, mesenchymal stem cells may be bone marrow, fat, umbilical cord blood-derived mesenchymal stem cells, and especially human-derived mesenchymal stem cells. In addition, the mesenchymal stem cells according to the present invention may be used to include not only stem cells, but also cultures of stem cells.

In the present invention, " culture product " may include a cell culture solution containing stem cells, a culture supernatant from which stem cells have been removed from the cell culture solution, and a dilution solution thereof. The composition of the culture may further include not only components necessary for culturing mesenchymal stem cells but also components that act synergistically with the proliferation of mesenchymal stem cells, . ≪ / RTI >

In the present invention, " cytokine " refers to an inflammatory cytokine. The inflammatory cytokine generally refers to a cytokine that induces an inflammatory reaction in the body. Examples of the cytokine include TNF-a, interferons (IFNs) , Interleukins (ILs), monocyte chemoattractant proteins, and the like. In particular, the cytokine according to the present invention may be, but is not limited to, interferon-gamma (IFN-γ), TNF-α, interferon-beta (IFN-β) and interleukin-10 (IL-10).

In the present invention, the cytokine may be selected from three or four combinations, and the combination may be a combination of IFN-y, TNF-a and IFN- ?, or IFN- ?, TNF- ?, and IL -10, or a combination of IFN-y, TNF-a, IFN-? And IL-10. The composition according to the present invention can enhance the ability of the mesenchymal stem cells to produce an inflammatory regulator by including the above-described combination of cytokines.

Accordingly, the present invention provides the use of three or four combined cytokines as described above for enhancing the inflammatory regulatory ability of mesenchymal stem cells.

In addition, in order to enhance the ability of the mesenchymal stem cells to produce an inflammatory regulator, the three or four cytokines thus combined are each treated at a concentration of 1 to 50 ng / ml, for example 1 to 30 ng / ml Lt; / RTI > Specifically, IFN-γ and TNF-α is a mesenchymal stem cell 1ⅹ10 ⁴ to 1ⅹ10 ⁶ cell number compared to each of 1 to 50 ng / ㎖ concentration of, for example, 1 to 30 ng / ㎖, 2 to 10 ng / ㎖, 3 And IFN-? And IL-10 may be contained at a concentration of 1 to 50 ng / ml, for example, 5 to 7 ng / ml, relative to the cell number of mesenchymal stem cells 1 x 10 ⁴ to 1 x 10 ⁶ , But are not limited to, 10 to 30 ng / ml, 15 to 25 ng / ml, 17 to 23 ng / ml, 18 to 22 ng / ml and 19 to 21 ng / .

In one embodiment, when combined with three cytokines, i.e. IFN-y, TNF- alpha and IFN-beta, or IFN-y, TNF- alpha and IL-10, May be contained in a ratio of 0.5 to 2: 0.5 to 2: 2 to 8, but not limited thereto, and may be, for example, in a ratio of 0.5 to 1: 0.5 to 1: 2 to 4, preferably 1: 1: .

In another embodiment, when combined with four cytokines, i.e. IFN-y, TNF-a, IFN-beta and IL-10, each cytokine is between 0.5 and 2: 0.5 to 2: To 8: 2 to 8, and is not limited thereto. For example, it may be contained in a ratio of 0.5 to 1: 0.5 to 1: 2 to 4: 2 to 4, preferably 1: 1: 4: 4 .

In addition, the cytokine may be added to the composition so that the total cytokine concentration is 4 to 80 ng / ml, such as 10 to 70 ng / ml, 15 to 65 ng / ml, 20 to 60 ng / .

According to the present invention, even when stimulating mesenchymal stem cells with a low concentration of each cytokine as described above, it is possible to enhance the ability of the mesenchymal stem cells to stimulate inflammation from the stem cells, Thereby maximizing the amount of regulatory factor produced.

In the present invention, the term "inflammatory modulator" refers to an inhibitor that inhibits inflammation or an immune response, and includes, for example, COX-2, PGE2 (prostaglandin E2; prostaglandin E2), interleukin- , TGF-beta, IDO (indoleamine 2,3-dioxygenase; indoleamine 2,3-deoxygenase), and the like. In the following examples, the effect of the three cytokines on enhancing the ability of the mesenchymal stem cells to regulate inflammation was confirmed. By measuring the expression levels of COX-2 and IDO as the inflammatory regulator, it was possible to confirm the ability of the mesenchymal stem cells to produce an enhanced inflammatory regulator in combination with the cytokine and at low treatment concentrations. Since COX-2 is known to be a key enzyme in the synthesis of PGE2, it can be predicted that the amount of PGE2 synthesized increases with the increase of COX-2 expression level. Therefore, since the synergistic effect according to the combination of the cytokines can be enhanced even at a low treatment concentration, the composition for enhancing the inflammatory stem cell production ability of the mesenchymal stem cells according to the present invention can provide It is possible to provide mesenchymal stem cells enhanced in the ability to regulate inflammatory factors for the prevention and treatment of immune diseases and inflammatory diseases.

Further, the present invention provides a method for producing a mesenchymal stem cell,

IFN-?;

TNF-a; And

Lt; RTI ID = 0.0 > IL-10, < / RTI > IFN-

Wherein the cytokine is treated with 1 to 50 ng / ml of the mesenchymal stem cell 1 × ^{10 4} to 1 × 10 ⁶ cells, respectively, to enhance the ability of the mesenchymal stem cells to produce an inflammatory regulator.

In the method for enhancing the ability of the mesenchymal stem cells to regulate inflammation, the combination of mesenchymal stem cells, cytokines, and treatment concentration can be applied or applied in the above-described manner.

The step of treating the mesenchymal stem cells with three or four cytokines may be an ex vivo treatment, wherein the mesenchymal stem cells comprise three or four cytokines in an amount of 1 to 50 ng / ml By culturing the mesenchymal stem cells in the medium for 12 hours to 48 hours, for example 12 hours to 36 hours, and 12 hours to 24 hours, the cytokine can be treated to the mesenchymal stem cells, but the treatment is not limited thereto .

In the present invention, three or four cytokines to be treated in mesenchymal stem cells may be treated at a concentration of 1 to 50 ng / ml, such as 1 to 30 ng / ml, respectively. Specifically, IFN-γ and TNF-α is a mesenchymal stem cell 1ⅹ10 ⁴ to 1ⅹ10 ⁶ cell number compared to each of 1 to 50 ng / ㎖ concentration of, for example, from 2 to 30 ng / ㎖, 3 to 10 ng / ㎖, 4 can be included in to 8 ng / ㎖ and, IFN-β and IL-10 is mesenchymal stem cells 1ⅹ10 ⁴ to cell number over each of 1 to 50 ng / ㎖ concentration of 1ⅹ10 ^6, for example 5 to 40 ng / ㎖, 10 To 30 ng / ml, and 15 to 25 ng / ml, but is not limited thereto.

In one embodiment, when combined with three cytokines, i.e. IFN-y, TNF- alpha and IFN-beta, or IFN-y, TNF- alpha and IL-10, May be contained in a ratio of 0.5 to 2: 0.5 to 2: 2 to 8, but not limited thereto, and may be, for example, in a ratio of 0.5 to 1: 0.5 to 1: 2 to 4, preferably 1: 1: .

In another embodiment, when combined with four cytokines, i.e. IFN-y, TNF-a, IFN-beta and IL-10, each cytokine is between 0.5 and 2: 0.5 to 2: To 8: 2 to 8, and is not limited thereto. For example, it may be contained in a ratio of 0.5 to 1: 0.5 to 1: 2 to 4: 2 to 4, preferably 1: 1: 4: 4 .

In addition, the cytokine may be added to the composition so that the total cytokine concentration is 4 to 80 ng / ml, such as 10 to 70 ng / ml, 15 to 65 ng / ml, 20 to 60 ng / . In the above method, the medium used for culturing mesenchymal stem cells may be a conventional medium known to be suitable for culturing stem cells in the art. For example, DMEM (Dulbecco's modified Eagle medium) or Keratinocyte-SFM Keratinocyte serum free medium may be used, but is not limited thereto. Other conditions such as temperature, culture time and the like can be followed by culturing conditions of conventional mesenchymal stem cells.

In the present invention, the medium may be supplemented with additives. (E. G., Serum, e. G., FBS, serum replacement, albumin, or essential amino acids and non-essential amino acids such as glutamine) in an isotonic solution. Furthermore, it has been found that lipid (fatty acid, cholesterol, serum HDL or LDL extract) and other components found in most types of storage medium of this kind (such as insulin or transferrin, nucleoside or nucleotide, pyruvate, G., Glucose, selenium, glucocorticoids such as hydrocortisone and / or reducing agents such as? -Mercaptoethanol).

In the following examples, IFN-? As a cytokine in mesenchymal stem cell culture; TNF-a; And IFN- [beta], IFN- [gamma]; TNF-a; And IL-10 and IFN-y; TNF-a; IFN-? And IL-10, the degree of inflammatory regulator expression of mesenchymal stem cells was confirmed by treatment with the cytokine. As a result, the cytokine treatment promoted the production of inflammatory regulators of mesenchymal stem cells. By treating the lower cytokine concentration, even when stimulated with a low concentration cytokine, the inflammation control of mesenchymal stem cells It was confirmed that the factor productivity increased. Therefore, it is possible to enhance and enhance the ability of the mesenchymal stem cells to produce an inflammatory regulator through the combination of the above cytokines and the low treatment concentration.

In addition, the present invention provides, as a cytokine,

IFN-?;

TNF-a; And

IFN-? And IL-10 as an active ingredient,

The cytokine was treated with 1 to 50 ng / ml of mesenchymal stem cells 1 x 10 ⁴ to 1 x 10 ⁶ cells,

There is provided a pharmaceutical composition for preventing or treating immune diseases and inflammatory diseases.

In the pharmaceutical composition of the present invention, the combination of mesenchymal stem cells, cytokines, the treatment concentration thereof, and the cytokine-treated mesenchymal stem cells can be applied or applied as described above.

In the present invention, the cytokine-treated mesenchymal stem cell refers to a mesenchymal stem cell having enhanced ability to produce an inflammatory regulator. As the mesenchymal stem cells are transplanted or administered to a subject, A therapeutic effect can be realized. In one embodiment, the cytokine-treated mesenchymal stem cells according to the present invention may be useful for the prevention and treatment of IDO or COX-2 mediated immune or inflammatory diseases. The IDO or COX-2-mediated immunological disease or inflammatory disease is an immune or inflammatory disease that may be caused by the fact that the IDO or COX-2 gene is not normally expressed on the immunity or inflammation-related mechanism or the protein is not secreted. it means.

In the present invention, " immunological diseases and inflammatory diseases " are collectively referred to as diseases in which inflammation is a major lesion and diseases in which a specific immune response occurs, such as autoimmune diseases, arthritis, bacterial infections, Inflammatory disease, < RTI ID = 0.0 > and / or < / RTI > The autoimmune disease may be selected from the group consisting of Crohn's disease, erythema disease, atopic dermatitis, rheumatoid arthritis, Hashimoto's thyroiditis, anemia, Edison's disease, type 1 diabetes, lupus, chronic fatigue syndrome, fibromyalgia, hypothyroidism, The present invention relates to the use of a compound of formula (I) for the manufacture of a medicament for the treatment of inflammatory bowel disease, inflammatory bowel disease, multiple sclerosis, myasthenia gravis, Meniere's syndrome, Guilian-Barre syndrome, Sjogren's syndrome, vitiligo, endometriosis, Asthma, and ulcerative colitis, but the present invention is not limited thereto.

In the pharmaceutical composition according to the present invention, the combination of the above cytokines enhances the ability of the mesenchymal stem cells to regulate inflammatory factors, thereby maximizing the amount of the inflammatory regulatory factor produced only by treatment with a low concentration of cytokine. Therefore, May have a therapeutic effect in patients with a disease or an inflammatory disease.

Accordingly, the present invention provides a therapeutic use of mesenchymal stem cells for the treatment of immune diseases and inflammatory diseases by enhancing the ability of mesenchymal stem cells treated with a combination of three or four cytokines at a low concentration, will be.

According to the following examples, the combination of the cytokine and the low treatment concentration can be treated at the time of culturing the mesenchymal stem cells to enhance the ability of the mesenchymal stem cells to produce the inflammatory regulator. Therefore, the combination of the cytokines proposed in the present invention can maximize the production of inflammatory regulatory factors of mesenchymal stem cells at a low treatment concentration by causing a synergistic effect, so that the treatment of the above diseases in patients exhibiting immunological diseases or inflammatory diseases Can be useful for.

In the present invention, the subject of the pharmaceutical composition may be a human or an animal in need of prevention and / or treatment of an immunological disease or an inflammatory disease.

The present invention also provides a method for treating an immunological disease or inflammatory disease comprising administering the cytokine-treated mesenchymal stem cells to a subject in need thereof.

As used herein, the term " subject " refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.

The pharmaceutical composition according to the present invention can be administered in a conventional manner via routes such as intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, and skin application.

The pharmaceutical compositions of the present invention may comprise a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not substantially stimulate the organism and does not interfere with the biological activity and properties of the administered ingredients. The pharmaceutically acceptable carrier in the present invention may be a mixture of saline, sterilized water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol and one or more components of these components, If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added to formulate into the form of an injection suitable for injection into tissues or organs. In addition, it may be formulated into an isotonic sterile solution or, in some cases, a dry preparation (especially a lyophilized preparation) which may be an injectable solution by adding sterile water or physiological saline. In addition, a target organ specific antibody or other ligand can be used in combination with the carrier so as to specifically act on the target organ.

Also preferably, the composition of the present invention may further include a filler, an excipient, a disintegrant, a binder, a lubricant, and the like. The composition of the present invention may also be formulated using methods known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal.

The term "administering" in the present invention means introducing the composition of the present invention to a patient by any suitable method, and the route of administration of the composition of the present invention may be carried out through various routes of oral or parenteral routes, ≪ / RTI > But are not limited to, intraperitoneal, intravenous, intramuscular, subcutaneous, intradermal, oral, topical, intranasal, intrathecal, rectal.

As used herein, the term "effective amount" means an amount required to delay or completely stop the onset or progression of a specific disease to be treated, and includes the amount of cytokine-treated mesenchymal stem cells contained in the pharmaceutical composition of the present invention An effective amount means an amount required to achieve the preventive or therapeutic effect of an immunological disease and an inflammatory disease. Thus, the effective amount may vary depending on factors such as the type of disease, the severity of the disease, the type and amount of other components contained in the composition, and the age, weight, general health status, sex and diet of the patient, ≪ RTI ID = 0.0 > and / or < / RTI > It will be apparent to those skilled in the art that the appropriate total daily dose may be determined by the practitioner within the scope of sound medical judgment.

For purposes of the present invention, the specific therapeutically effective amount for a particular patient will depend upon a variety of factors, including the type and extent of the response to be achieved, the specific composition including whether or not other agents are used, the age, weight, And various factors including diet, time of administration, route of administration and minute of composition, duration of treatment, drugs used or co-used with the specific composition, and similar factors well known in the medical field.

As used herein, " treatment " means an approach for obtaining a beneficial or desired clinical result. For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, reduction in the extent of disease, stabilization (i.e., not worsening) of the disease state, (Either partially or totally), detectable or undetected, whether or not an improvement or temporary relief or reduction Also, " treatment " may mean increasing the survival rate compared to the expected survival rate when not receiving treatment. &Quot; Treatment " refers to both therapeutic treatment and prophylactic or preventative measures. Such treatments include treatments required for disorders that have already occurred as well as disorders to be prevented. &Quot; Relaxing " disease means that the extent of the disease state and / or undesirable clinical symptoms are reduced and / or the time course of the progress is slowed or lengthened, do.

Advantages and features of the present invention and methods of achieving them will become apparent with reference to the embodiments described in detail below. The present invention may, however, be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. Is provided to fully convey the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.

The present invention relates to a method for enhancing the inflammatory regulator-producing ability of mesenchymal stem cells treated with cytokine, cytokine-treated mesenchymal stem cells, and mesenchymal stem cells, The present invention relates to a composition for preventing and treating immunological diseases and inflammatory diseases. More specifically, the present invention provides mesenchymal stem cells having enhanced ability to produce inflammatory factors by treating three or four cytokines at a low concentration during mesenchymal stem cell culture , COX-2, IDO and the like can be maximized to produce therapeutic effects on immune diseases and inflammatory diseases.

Fig. 1 shows the results of confirming the expression level of IDO according to the cytokine combination.
2 shows the result of confirming the expression level of COX2 according to cytokine combination.

Hereinafter, the present invention will be described in detail with reference to examples. The following examples illustrate the present invention, but the scope of the present invention is not limited to the following examples.

Examples. Cultivation of cytokine-treated mesenchymal stem cells

Human bone marrow-derived mesenchymal stem cells were seeded in 12 wells at 1 × 10 ⁵ cells, stimulated by combination of IFN-γ, TNF-α, IFN-β and IL-10, And cultured to obtain cytokine-treated mesenchymal stem cells. At this time, IFN-γ and TNF-α were stimulated by 5 ng / ml for IFN-β and 20 ng / ㎖ for IL-10, respectively.

Experimental example. Confirmation of expression level of inflammatory regulator of mesenchymal stem cells (Western blot)

The various combinations of mesenchymal stem cells obtained in the above Examples and Comparative Examples were subjected to disruption using 1% Nonidet-P40 buffer supplemented with 2 mM dithiothreitol and a protease inhibitor cocktail (Roche, US) lysate. Cell lysate was resolved by 12% SDS-PAGE, transferred to nitrocellulose membrane and immunostained with primary antibody (IDO, COX-2, β-actin). After immunostaining with secondary antibody, the protein was detected by ECL (enhanced chemiluminescence). The results are shown in Fig. 1 (IDO production amount) and 2 (COX-2 production amount).

1 and 2, it was confirmed that the group treated with only two cytokines (IFN-γ and TNF-α) showed a higher level of IDO and COX2 production than the untreated group. In addition, the three combination groups in which the cytokines were combined with the two cytokines were higher in IDO and COX2 than in the two combinations except for the IDO production of the IFN-? Respectively. In particular, it was confirmed that the synergistic effect maximizes the production of IDO and COX2 in the experimental group combining the four cytokines.

(IFN- ?, TNF- ?, IFN-? Or IL-10) or four cytokines (IFN- ?, TNF- ?, IFN- ?, and IL -10), the expression of IDO and COX-2, which are inflammatory factors, was maximized in the case of stimulation of mesenchymal stem cells. It is expected to be applicable.

Claims (13)

A composition for enhancing an inflammatory regulator-producing ability of mesenchymal stem cells comprising IFN-y, TNF-alpha, IFN-beta and IL-10 as cytokines,
Each of the cytokines comprises 1 to 50 ng / ml of mesenchymal stem cells 1 x 10 ⁴ to 1 x 10 ⁶ cells, and the IFN- ?, TNF- ?, IFN- ?, and IL-10 are in the range of 0.5 to 2: 0.5 To about 2: 2 to about 8: 2 to about 8: 1. The method according to claim 1,
Wherein the mesenchymal stem cells are fat, bone marrow, or umbilical cord blood-derived stem cells. delete The method according to claim 1,
Inflammatory regulators are composed of prostaglandin E2, PGE2, indoleamine 2,3-dioxygenase (IDO), interleukin-10 (IL-10) and TGF- ≪ / RTI > Culturing the mesenchymal stem cells by treating with IFN-y, TNF-a, IFN-? And IL-10 as cytokines,
Wherein the cytokine is treated with 1 to 50 ng / ml of mesenchymal stem cells 1 x 10 ⁴ to 1 x 10 ⁶ cells, and the IFN- ?, TNF- ?, IFN-? And IL-10 are treated with 0.5 to 2: 0.5 To about 2: 2 to about 8: 2 to about 8: < RTI ID = 0.0 > 2. ≪ / RTI > 6. The method of claim 5,
The mesenchymal stem cell is a method for enhancing the ability of mesenchymal stem cells, which are stem cells derived from fat, bone marrow or cord blood, to produce an inflammatory regulator. delete 6. The method of claim 5,
Wherein the step of treating and culturing mesenchymal stem cells with cytokines is an ex vivo treatment. 6. The method of claim 5,
Inflammatory regulators are composed of prostaglandin E2, PGE2, indoleamine 2,3-dioxygenase (IDO), interleukin-10 (IL-10) and TGF- Wherein the at least one protein is selected from the group consisting of: < RTI ID = 0.0 > Mesenchymal stem cells treated with IFN-?, TNF- ?, IFN-? And IL-10 as cytokines as an active ingredient,
Wherein the cytokine is treated with 1 to 50 ng / ml of mesenchymal stem cells 1 x 10 ⁴ to 1 x 10 ⁶ cells, and the IFN- ?, TNF- ?, IFN-? And IL-10 are treated with 0.5 to 2: 0.5 To about 2: 2 to about 8: 2 to about 8: < RTI ID = 0.0 > 1. ≪ / RTI > delete delete 11. The method of claim 10,
Autoimmune diseases include autoimmune diseases such as Crohn's disease, erythema, atopic dermatitis, rheumatoid arthritis, Hashimoto's thyroiditis, anemia, Edison's disease, type 1 diabetes, lupus, chronic fatigue syndrome, fibromyalgia, hypothyroidism and hyperlipemia, scleroderma, Inflammatory bowel disease, multiple sclerosis, myasthenia gravis, Meniere's syndrome, Guilian-Barre syndrome, Sjogren's syndrome, vitiligo, endometriosis, psoriasis, vitiligo, systemic scleroderma, asthma And ulcerative colitis. ≪ RTI ID = 0.0 > 21. < / RTI >

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