KR101583666B1 - Composition for preventing or treating autoimmune diseases or transplant rejection comprising human fetal cartilage-derived cells - Google Patents

Abstract

The present invention relates to a composition for preventing or treating immune diseases using a fetal cartilage-derived cell capable of suppressing / regulating an immune response occurring during tissue transplantation. More particularly, the present invention relates to a composition for preventing or treating immune diseases, Inhibits the proliferation of activated lymphocytes and controls the expression of the immunocytokines produced by the activated lymphocytes. The proliferation ability and the ability to differentiate into various cell tissues are superior to those of adult stem cells, And can be usefully used as an immunomodulator that can regulate an immune response that may occur when transplanting together.

Description

[0001] The present invention relates to a composition for preventing or treating autoimmune diseases or transplant rejection diseases using fetal cartilage-derived cells,

The present invention relates to a novel immunomodulating composition for regenerating damaged tissue, and more particularly, to a composition for preventing and treating immunological diseases using fetal cartilage-derived cells having superior proliferative power or cell phenotype maintenance than known adult stem cells.

Recently, transplantation therapy using stem cell and tissue engineering has been actively developed as a treatment method for intractable tissue injury. However, there is a problem that it is difficult to develop a stable cell therapy agent that does not cause the immune response of the human body because of the excellent therapeutic effect with such transplantation therapy technique. While many attempts have been made to develop adult stem cells and use them as cell therapy agents, only immune responses have been found, but identification of immunomodulatory ability has yet to be confirmed.

Adult stem cells are stem cells capable of differentiating into the ground, cartilage, bone, muscle cells, etc., and are highly proliferated and highly valuable for tissue regeneration. It has been reported that specific adult stem cells such as mesenchymal stem cells do not develop an immune response by itself and have immunosuppressive ability (see Placenta-Derived Multipotent Cells Exhibit Immunosuppressive Properties That Interference- , STEM CELLS, 2006), but there is no report yet on stem cells with immunomodulating ability.

Immunomodulators, known as drugs to enhance the immune function of living organisms, have been proven to have impaired immune response at various stages. Based on the immunological background of cancer patients, immune stimulating agents have been used to improve immune function in cancer patients. RTI ID = 0.0 > immunotherapy, < / RTI > When live cells are used, cell wall skeleton (CWS) is often used as a purified cell component because of its stability and side effects. Immunomodulators are mainly used for the purpose of cancer treatment.

Therefore, it is very important to develop a desirable immunomodulator that can selectively control unnecessary immune responses in the human body when transplantation therapy using stem cell and tissue engineering is performed.

It is an object of the present invention to provide a composition for preventing or treating immunological diseases using cells derived from fetal cartilage capable of inhibiting the proliferation of activated lymphocytes without causing an immune response.

In order to achieve the above object, the present inventors have found that cells isolated from fetal cartilage tissue can overcome the limitations of existing stem cells by regulating damaged tissues and regulating immune responses that may occur during transplantation, The present inventors have developed an immunomodulating composition which can be used as an immunomodulating agent.

The present invention provides a composition for preventing or treating immune diseases comprising cells derived from fetal cartilage as an active ingredient.

The fetal cartilage-derived cells can express surface antigen proteins of stem cells. In one embodiment, the fetal cartilage-derived cells express CD29, CD44, CD90, and CD105, which are bone marrow-derived mesenchymal stem cell- , But SSEA4 and Oct-4, which are specific surface antigen of embryonic stem cells, can be expressed.

The fetal cartilage-derived cells can control the expression of the immunocytokines produced in activated lymphocytes.

More specifically, the fetal cartilage-derived cells can inhibit the expression of TNF-a, IL-13, and IFN-y associated with an immune response or an inflammatory reaction. In addition, the fetal cartilage-derived cells can control the immune activity by not affecting or increasing the expression of IL-10, an anti-inflammatory cytokine.

The immunomodulating composition may comprise fetal cartilage-derived cell extracts or secreted molecules.

The fetal cartilage-derived cells are prepared by: (a) separating and culturing chondrocytes in fetal cartilage tissue; (b) isolating cells expressing the surface antigen protein of the stem cells in the cells isolated and cultured in the step (a); And (c) confirming the proliferative capacity and pluripotency of the cells isolated in the step (b), thereby verifying the fetal cartilage-derived stem cells.

Immunomodulation of the fetal cartilage-derived cells is carried out by (a) analyzing whether the cells isolated from fetal cartilage tissue and lymphocytes isolated from allogeneic whole blood are cultured together to proliferate lymphocytes, (b) interferon known as immunointerferon A method of analyzing the characteristics of cells separated from fetal cartilage tissue reacted with gamma, or (c) a method of identifying an immunomodulatory cytokine secreted upon culturing in step (a) .

The fetal cartilage-derived cells according to the present invention do not cause an immune response but rather inhibit the proliferation of activated lymphocytes and control the expression of the immunocytokines produced in the activated lymphocytes. The proliferative ability and various cell tissues Can be used as an immunomodulator capable of regulating the immune response that may occur during transplantation as well as the regeneration of injured tissue.

FIG. 1 shows that fetal cartilage-derived cells from the proliferation inhibited cells and the lymphocytes isolated from the blood of the same kind were mixed and cultured, and the proliferation of the lymphocytes was observed, indicating that the fetal cartilage-derived cells did not induce an immune response.
FIG. 2 is a graph showing the results of analysis of lymphocyte proliferation after treatment of mixed lymphocyte reaction with ConA in fetal cartilage-derived cells and adult chondrocytes suppressing proliferation, respectively. As a result, fetal cartilage-derived cells inhibited the activation of lymphocytes by concanavalin A. .
FIG. 3 shows that immunogenicity of fetal cartilage-derived cells according to presence or absence of interferon stimulation was analyzed by a flow cytometry analyzer to show that the fetal cartilage-derived cells were not activated by IFN-γ.
FIG. 4 shows that fetal cartilage-derived cells and progenitor cartilage cells treated with concha or valine A, respectively, regulate the expression of immunocytokines, which are expressed in lymphocytes. will be.
Fig. 5 shows the inflammation-relieving effect of fetal cartilage-derived cells using an animal model.

Hereinafter, the present invention will be described in more detail.

The present invention provides a composition for preventing or treating immune diseases using the cells derived from fetal cartilage tissue as an active ingredient which can overcome the limitations of existing stem cells by regulating the immune response that may occur during transplantation with regeneration of damaged tissue do.

In other words, the cells were isolated from the fetal cartilage tissue and reacted with the same type of lymphocytes to analyze the immune response. As a result, it was confirmed that the immune response was not caused, and thus the composition for preventing or treating immune diseases clinically applicable to provide.

The fetal cartilage-derived cells can be obtained by (a) confirming the proliferation of lymphocytes in the cells when the cells isolated from the fetal cartilage tissue and the lymphocytes isolated from the allogeneic blood are co-cultured, or (b) reacting with interferon gamma known as immunointerferon (C) confirming the immunoregulatory cytokine secreted upon culturing in step (a). However, in the case of cells derived from fetal cartilage tissue-derived cells Can be analyzed by conventional methods known to those skilled in the art, and thus the present invention is not limited thereto.

The fetal cartilage-derived cells express SSEA4 and Oct-4, which are specific surface antigen of embryonic stem cells, while expressing CD29, CD44, CD90 and CD105, which are bone marrow-derived mesenchymal stem cell-specific surface antigen in adults.

The fetal cartilage-derived cells are prepared by: (a) separating and culturing chondrocytes in fetal cartilage tissue; (b) isolating cells expressing the surface antigen protein of the stem cells in the cells isolated and cultured in the step (a); And (c) confirming the proliferative capacity and pluripotency of the cells isolated in the step (b), thereby verifying the fetal cartilage-derived stem cells.

The chondrocytes isolated from the fetal cartilage tissue can be obtained from 12-15 week-aborted fetuses and obtained by using collagenase or trituration.

The fetal cartilage-derived cells can express surface antigen proteins of stem cells. In one embodiment, the fetal cartilage-derived cells express CD29, CD44, CD90, and CD105, which are bone marrow-derived mesenchymal stem cell- , But SSEA4 and Oct-4, which are specific surface antigen of embryonic stem cells, can be expressed.

The fetal cartilage-derived cells can control the expression of the immunocytokines produced in activated lymphocytes. More specifically, the fetal cartilage-derived cells can inhibit the expression of TNF-a, IL-13, and IFN-y associated with an immune response or an inflammatory reaction. In addition, the fetal cartilage-derived cells can control the immune activity by not affecting or increasing the expression of IL-10, an anti-inflammatory cytokine.

As an example, simultaneous expression of surface antigens of mesenchymal stem cells and surface antigens of embryonic stem cells in the fetal cartilage-derived cells can be performed using Fluorescent-Assisted Cell Sorting (FACS) and Magnetic-Activated Cell Sorting (MACS) method.

The fetal cartilage-derived cells include musculoskeletal tissue cells including cartilage, bone, muscle, skin and fat cells; Nerve cells; Hepatocytes; Pancreatic cells; Myocardial cells; And vascular cells, as described above.

Since the known immunomodulating agents are mainly used for cancer treatment, there is a limit in that they can not play a role of a therapeutic agent for tissue transplantation or tissue regeneration. The fetal cartilage-derived cells of the present invention are more likely to be used for cell therapy than adult stem cells It has excellent proliferation and differentiation potential, and has the ability to regulate the immune response without causing an immune response in the same species. That is, according to one embodiment, the cells of the fetal cartilage do not stimulate lymphocytes of the same type, but when the lymphocyte activation is induced, the proliferation of lymphocytes is decreased, and in particular, the TNF-a, IL-13 , But the immunoreactivity is regulated by inhibiting or increasing the expression of IL-10, which is known as an anti-inflammatory cytokine, but not IFN-γ.

The composition for immunomodulation according to the present invention includes, but is not limited to, a cell therapy agent, and can be utilized in various therapeutic agent forms. Specifically, fetal cartilage-derived cell extracts or secreted molecules. Cell extracts or secreted cells derived from fetal cartilage may be of any type that can be obtained from fetal cartilage derived cells and may be used not only as a cell itself but also as a suspension or a culture thereof or a concentrate thereof, A supernatant, a dried product, a diluent, or any product produced through metabolism of the cell.

The immunological diseases may be autoimmune diseases, inflammatory diseases, and graft-rejection diseases of cells, tissues or organs, and more particularly, arthritis, autoimmune hepatitis, rheumatoid arthritis, osteoarthritis, insulin dependent diabetes, ulcerative colitis, But are not limited to, multiple sclerosis, autoimmune myocarditis, scleroderma, severe muscular atrophy, multiple myositis / dermatomyositis, Hashimoto's disease, autoimmune cytopenias, Sjogren's syndrome, vasculitis syndrome and systemic lupus erythematosus.

Compositions for the prevention or treatment of immune diseases in one embodiment of the present invention can be prepared in various ways by mixing with a pharmaceutically acceptable carrier. For example, a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersant, a stabilizer, a suspending agent, a coloring matter, a perfume or the like may be mixed and used as a pharmaceutically acceptable carrier. In the case of injections, a buffering agent, a preservative, an anhydrous agent, a solubilizing agent, an isotonic agent, a stabilizer and the like may be mixed and used. In the case of injections, they may be formulated in unit dosage ampoules or in multi-dose dosage forms. In topical administration, a base, an excipient, a lubricant, a preservative, etc. may be used. However, the present invention is not limited thereto. Those skilled in the art can formulate compositions for preventing or treating immune diseases of the present invention in various forms, and may include various carriers and additives. I will understand.

The composition of the present invention can be administered in various routes to mammals such as rats, mice, livestock, and humans, depending on the severity of the disease. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections. The dosage of the active ingredient in the composition may vary depending on the route of administration, the severity of the disease, the age, gender, body weight, etc., and may be administered once to several times per day.

In the present invention, "stem cells" are cells having the ability to differentiate into various kinds of body tissues, that is, undifferentiated cells. The stem cells include embryonic stem cells (pluripotent stem cells) that can be made using human embryos and adult stem cells (multipotent stem cells) such as bone marrow cells that constantly make blood cells .

In the present invention, the term "embryonic stem cell" refers to a cell of an embryonic stem that has undergone modification for 14 days, and can be differentiated into all the cells and tissues that constitute the human body. Lt; / RTI > An embryo forms a blastocyst through cell division, inside which there is a cell mass called an inner cell mass, which forms an embryo. When the cells of the inner cell mass are separated from the blastocyst and cultured, the differentiation does not occur but the embryonic stem cell still has differentiation ability.

In the present invention, "Adult Stem Cell" is extracted from cord blood and blood of a cord blood (umbilical cord blood) or a multiparous adult. It is a raw material just before being differentiated into specific organs such as bone, liver, Cells. These include hematopoietic stem cells and Mesenchymal Stem Cells, neural stem cells, which are widely used as materials for regenerative medicine. In addition, there are stem cells in the liver, epidermis, and pancreas. The adult stem cells are difficult to proliferate and tend to differentiate easily. Instead of using a variety of adult stem cells, they can be used for long-term regeneration required in actual medicine, .

In the present invention, the term "mesenchymal stem cell (MSC)" refers to a bone, a cartilage, a fat, a tendon, a nerve tissue, a fibroblast ) And muscle cells (muscle cells), such as specific organs before the differentiation of cells is called a multipotent progenitor (multipotent progenitor).

In the present invention, the term "cell therapeutic agent" is a medicament (US FDA regulation) used for the purpose of treatment, diagnosis and prevention of cells and tissues prepared by separation, culture and special works from human, Refers to a drug used for therapeutic, diagnostic and prophylactic purposes through a series of actions such as alive, homologous, or xenogeneic cell propagation, screening, or otherwise altering the biological characteristics of a cell to restore it. The cell therapy agent is classified into a somatic cell therapy agent and a stem cell treatment agent depending on the degree of cell differentiation, and the present invention particularly relates to a stem cell therapeutic agent.

Hereinafter, the present invention will be described in more detail based on examples. It should be understood that the following embodiments of the present invention are only for embodying the present invention and do not limit or limit the scope of the present invention. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. The references cited in the present invention are incorporated herein by reference.

Example  One. Fetal cartilage origin  Identification of lymphocyte response by cells

(IRB) for the isolation and culture of fetal cartilage-derived cells and human peripheral blood-derived lymphocytes. The fetal cartilage-derived cells were obtained from Sanbon Jeil Hospital or Ajou University Hospital for consent at the time of surgery Human fetal cartilage was collected, and human peripheral blood was collected from healthy donors after the donor recruitment announcement, and lymphocytes were isolated.

Methods for the isolation and culturing of fetal cartilage-derived cells were carried out as described in detail in the previously filed patent application No. 2013-0090233 [Fetal cartilage tissue-derived stem cell source and cell therapy agent containing the same]. In other words, the collected cartilage tissue was washed several times with phosphate buffered saline (PBS), finely cut, and cultured in a culture medium (DMEM) containing 0.1% collagenase and 1% fetal bovine serum (FBS) And the cells were allowed to stand for 12 hours in an incubator at 37 ° C and 5% CO ₂ . Cells isolated by collagenase were washed with a culture medium containing 10% FBS. The cells suspended in the culture medium were centrifuged at 1700 rpm for 10 minutes, the supernatant was removed, and the collected cells were collected and suspended again in the culture medium. At this time, some cells were stained with trypan blue, and cell numbers were measured using a hemocyte plate under an optical microscope. The cells suspended in the culture medium were placed in a culture container at a concentration of 8 × 10 ⁴ / cm ² and then cultured in an incubator at 37 ° C. and 5% CO ₂ . Subculture was done when the cells filled 80 to 90% of the culture vessels.

As described in Patent Application No. 2013-0090233, the obtained fetal cartilage-derived cells were analyzed by using a flow cytometer (FACScanto II), and found that CD29 (Integrin β1 chain), CD44 (4) and (4), which are specific markers of embryo-derived stem cells, as well as high levels of HA receptor, CD90 (Thy-1) and CD105 (Endoglin).

In addition, the peripheral blood-derived lymphocytes were separated from the whole blood using Ficoll.

Two cells were cultured together to confirm lymphocyte reaction by fetal cartilage - derived cells. At this time, the fetal cartilage-derived cells were treated with gamma rays (3000 rad, 5 minutes) to inhibit proliferation, and the cells were divided into 10 ² , 10 ³ and 10 ⁴ cells in a 96-well plate. Cells derived from fetal cartilage After 16 hours, lymphocytes isolated from allogeneic blood were plated on the same plate at 10 ^{5 and} cultured together for 4 days. At this time, the cell division ratios of the fetal cartilage-derived cells and the lymphocytes were 1: 1000, 1: 100, and 1:10, respectively. To confirm the normal activity of the isolated lymphocytes, 10 μg / ml of concanavalin A (T cell mitogen) was added to induce the activity of the lymphocyte-only group for 4 days. After that, the floating lymphocytes and supernatant were collected and subjected to the BrdU assay (Cell proliferation ELISA, BrdU, Roche Diagnostice, Germany).

Lymphocytes treated with concanavalin A showed increased proliferation over untreated lymphocytes. This indicates that the isolated lymphocytes were normally activated. And that the group treated with fetal cartilage-derived cells did not proliferate (see FIG. 1). It was judged that the cells did not activate lymphocytes by fetal cartilage - derived cells.

Example  2. Fetal cartilage origin  By cell Derived from peripheral blood  Reduction / inhibition of proliferation of lymphocytes

(IRB) for the isolation and culturing of fetal cartilage-derived cells, adult chondrocytes and human peripheral blood-derived lymphocytes. The fetal cartilage-derived cells were obtained from Sanbon Jeil Hospital or Ajou University Hospital, After receiving consent, fetal cartilage was collected. Adult chondrocytes were obtained from the patients who underwent artificial cartilage replacement in the orthopedic department of Ajou University Hospital. Human peripheral blood was collected from healthy donors after the donor recruitment announcement, and lymphocytes were isolated.

Separation and culture of fetal cartilage-derived cells were carried out in the same manner as in the previous example. In order to separate peripheral blood-derived lymphocytes, Ficoll was used to generally separate lymphocytes from whole blood.

Mixed lymphocyte reaction was performed as a method to confirm the inhibition of lymphocyte proliferation of fetal cartilage - derived cells. At this time, the fetal cartilage-derived cells were treated with gamma rays (3000 rad, 5 minutes) to inhibit proliferation, and the cells were divided into 10 ³ and 10 ⁴ cells in a 96-well plate. Fetal cartilage-derived cells were cultured 16 hours after the injection of 10 ⁵ lymphocytes isolated from allogeneic blood on the same plate for 4 days. At this time, the cell division ratio of the fetal cartilage-derived cells and the lymphocytes was 1: 100 and 1:10. And 10 μg / ml of concha or valine A (T cell mitogen) was added for 4 days to induce lymphocyte activity. After that, the floating lymphocytes and supernatant were collected and subjected to the BrdU assay (Cell proliferation ELISA, BrdU, Roche Diagnostice, Germany). In addition, the test was performed using the same method for adult chondrocytes used as the comparative group.

Lymphocytes treated with concanavalin A showed increased proliferation over untreated lymphocytes. In the group treated with fetal cartilage-derived cells, the proliferation of lymphocytes was decreased as the number of fetal cartilage-derived cells increased (see FIG. 2). It was concluded that fetal cartilage - derived cells have the property of inhibiting the proliferation of concha or valine A - activated lymphocytes. On the other hand, adult chondrocytes showed activated lymphocyte proliferation. It can be seen that adult chondrocytes are not capable of inhibiting lymphocyte proliferation.

Example  3. Fetal cartilage origin  Identification of cell immunogenicity

After culturing for 7 days with 200 units / ml of interferon gamma (IFN-γ) known as immunointerferon, the cell-mediated immune response was induced in the fetal cartilage-derived cells and the characteristics of the fetal cartilage-derived cell surface were confirmed by flow cytometry .

The leukocyte antigen (HLA) ABC and DR, which are known to cause severe rejection when organ organs enter the body when they are transplanted into a type 1 leukocyte, . As a result, fetal cartilage-derived cells showed that no human leukocyte antigen appeared regardless of interferon stimulation (see FIG. 3).

In addition, we analyzed whether CD80, CD86 (B7.1, B7.2), which is a co-stimulatory molecule that is activated upon induction of signal transduction from the immune response to antigen-presenting cell T cells, (See FIG. 3).

Example  4. Fetal cartilage origin  Cell Cytokine  produce

Previous results showed that the fetal cartilage - derived cells did not have an antigen capable of causing an immune response and inhibited the proliferation of allogeneic lymphocytes. In the next experiment, mixed lymphocyte reaction was performed to confirm the cytokine production of fetal cartilage-derived cells. At this time, the fetal cartilage-derived cells were treated with gamma rays (3000 rad, 5 minutes) to inhibit proliferation and were divided into 10 ⁵ and 10 ⁶ cells in a 12-well plate. After 16 hours, the separated lymphocytes were divided into 10 ⁶ parts. At this time, the cell division ratio of the fetal cartilage-derived cells and the lymphocytes was 1:10 and 1: 1. And 10 μg / ml of concha or valine A (T cell mitogen) was added for 4 days to induce lymphocyte activity. As a comparative group, allogeneic lymphocytes, lymphocytes treated with concanavalin A, and fetal cartilage derived cells were cultured. After 4 days of culture, the supernatant was collected, centrifuged at 2000 rpm for 5 minutes to remove floating lymphocytes, and the supernatant was collected again and subjected to cytokine-ELISA analysis (eBioscience, Ready-SET-Go ELISA set).

Tumor necrosis factor-alpha (TNF-α), a cytokine that appears in acute immune or inflammatory responses, was produced in the lymphocyte group activated by concanavalin A, but not in the cultured cells of fetal cartilage-derived cells Group (Fig. 4 (A)).

In addition, interleukin-13 (IL-13), a lymphocyte produced in activated helper T cells, and interferon-gamma produced in T cells by inflammatory and immunostimulatory responses, gamma, IFN-y), the production was decreased when the ratio of the number of fetal cartilage-derived cells to the number of lymphocytes was 1: 1 compared to 1:10 (see FIGS. 4 (B) and 4 ).

Interleukin-10 (IL-10), which is produced by regulatory T cells or dendritic cells, is a cytokine that is used as an anti-inflammatory agent by inducing immune tolerance by inhibiting immune cells that attack self- As shown in FIG. 4 (D), when the fetal cartilage-derived cells and lymphocytes were reacted, the production of interleukin-10 was increased as the proportion of fetal cartilage-derived cells increased.

When the cells isolated from adult bone marrow were reacted with lymphocytes in the same manner, it was confirmed that interferon-gamma production was reduced with the same tendency as in fetal cartilage-derived cells (see FIG. 4 (E)). However, the production of interleukin-10, which acts on immune tolerance, decreased as the number of cells increased (see Fig. 4 (F)).

Example  5. Fetal cartilage origin  Inflammatory edema mitigation effect of cells

Tumor necrosis factor-alpha (TNF), which is an inflammatory factor, is increased by interleukin-10 (IL-10) and anti-inflammatory drugs -α) and interferon-gamma (IFN-γ) were decreased. In the next experiment, the effect of inflammation on fetal cartilage-derived cells was confirmed using an animal model. A complete Freund's adjuvant (CFA), a substance that induces a cell-mediated immune response in the rat knee joint cavity, was injected with 100 μl of 100 μl of insulin syringe to induce inflammation. Four days after the injection of the complete Freund's adjuvant, 5 × 10 ⁶ cells of fetal cartilage-derived cells were made into 50 μl and injected at the same position using an insulin syringe. As a comparative group, normal rats without induction of inflammation, complete Freund's adjuvant, and rats injected with phosphate buffered saline (PBS) instead of fetal cartilage-derived cells were injected with complete Freund's adjuvant to induce inflammation, And rats injected with 0.5 mg of triamcinolone, which is a therapeutic agent for inflammation used in clinical practice after induction of inflammation, were used. At this time, all groups injected the same amount. After that, the knee circumference of the rats was measured every day and the degree of edema was observed.

On the 4th day after injection of the complete Freund's adjuvant, the peripheral area of the rats was measured, and it was confirmed that the peripheral area increased and edema occurred compared with the normal rats. Triamcinolone, which is injected intra-articularly for the relief of inflammation, was a steroid agent, and it was confirmed that the rats in the group treated with this treatment had decreased edema due to reduction of the knee circumference from the 2nd day after the drug treatment (see FIG. 5).

In rats injected with fetal cartilage-derived cells having immunomodulating ability, edema was reduced as the knee circumference decreased from day 7 (see FIG. 5).

Claims (9)

Derived cells expressing bone marrow-derived mesenchymal stem cell-specific surface antigens CD29, CD44, CD90 and CD105, and SSEA4 and Oct-4 expressing embryonic stem cell-specific surface antigens, A pharmaceutical composition for preventing or treating a disease or a graft-rejection disease of a cell, tissue or organ. delete delete [Claim 2] The method according to claim 1, wherein the fetal cartilage-
(a) separating and culturing chondrocytes in fetal cartilage tissue;
(b) separating mesenchymal stem cells and cells expressing the surface antigen protein of embryonic stem cells simultaneously in the cells isolated and cultured in step (a); And
(c) a step of obtaining a stem cell source derived from a fetal cartilage tissue by verifying the proliferative capacity and pluripotentiality of the cells isolated in the step (b), thereby producing an autoimmune disease or a cell, tissue or organ A pharmaceutical composition for the prevention or treatment of graft rejection disease. [Claim 2] The pharmaceutical composition according to claim 1, wherein the fetal cartilage-derived cell regulates the expression of an immunocytokine produced in an activated lymphocyte, or a prophylactic or therapeutic agent for a transplant rejection disease of a cell, tissue or organ. The method according to claim 1, wherein the fetal cartilage-derived cell inhibits the expression of TNF-a, IL-13 or IFN-y associated with the immune response. Or < / RTI > [Claim 2] The method according to claim 1, wherein the fetal cartilage-derived cell does not affect the expression of IL-10, an anti-inflammatory cytokine, or modulates immunological activity by increasing expression of IL-10. A pharmaceutical composition for preventing or treating transplant rejection diseases of an organ. The pharmaceutical composition according to claim 1, wherein the composition comprises fetal cartilage-derived cell extract or secretion, or a pharmaceutical composition for preventing or treating an autoimmune disease or a cell, tissue or organ transplant rejection disease. The use according to claim 1, wherein the autoimmune disease or cell, tissue or organ transplant rejection disease is selected from the group consisting of insulin dependent diabetes mellitus, Crohn's disease, multiple sclerosis, scleroderma, severe muscle weakness, Hashimoto's disease, autoimmune cytopenias, Sjogren's syndrome, Lupus erythematosus, and lupus erythematosus. 2. A pharmaceutical composition for preventing or treating autoimmune disease or a cell, tissue or organ transplant rejection disease.

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