TR201902008A2 - A NEW GENERATION OF PANCREATIC TISSUE AND PRODUCTION METHOD THAT CAN AVOID AUTOIMMUNE ATTACKS AND INCREASE INSULIN - Google Patents

Abstract

The invention relates to a therapeutic method that enables the reprogramming of the immune system that causes autoimmunity in T1D (Type 1 Diabetes) and the production of glucose sensitive insulin secreting tissues by developing new techniques in endocrine pancreatic engineering. In the invention, in the tissue production method that synthesizes new generation insulin, a hybrid suppression technique is developed by using two separate areas of tissue engineering, decellularization and 3D bioprinting techniques, and a bioactive matrix is produced by using stem cells of the pancreas's own extracellular matrix. By differentiation of autologous mesenchymal stem cells, insulin-producing beta cells are obtained and insulin-secreting endocrine pancreas-like functional tissue part is produced. In the present invention, as the therapeutic method for the treatment of T1D, first of all, the autoimmune immune system attacking insulin-producing β-cells in T1D is suppressed. After autoimmune attacks are prevented in individuals with T1D, the insulin-producing tissue piece created within the scope of the invention is transplanted to individuals with T1D in the form of subcutaneous biochips. Thus, this endocrine pancreatic-like tissue, expressed as a biochip, can bring blood sugar to a healthy level by secreting insulin in response to glucose in diabetic individuals.

Description

DESCRIPTION 0NEW GENERATION PANCREATIC TISSUE AND PRODUCTION METHOD THAT CAN AVOID TOIMMUNE ATTACKS AND SECRET INSULIN Technical Field Buluş, TlD (A therapeutic method that enables the reprogramming of the immune system that causes autoimmunity in Type 1 Diabetes and the development of new techniques in endocrine pancreas engineering, the production of glucose-sensitive insulin-secreting tissues relates to production. Background Art Type 1 diabetes is an autoimmune disease and the death of beta cells in the body or their destruction to the point of not producing insulin is due to the autoimmunity involved in the pathogenesis of the disease [1]. Type 1 diabetes mellitus (TlD), which is a disease characterized by insulin production or complete inability to produce insulin, is one of the diseases for which intensive efforts are currently being made to treat, and the discovery of insulin has been an important development for diabetic patients. Insulin, which cannot be produced sufficiently by the pancreas, has begun to be supplied exogenously. For this reason, TlDM is also called insulin-dependent diabetes mellitus (IDDM). Although TDM patients can live their lives thanks to the insulin they receive externally through injection, low quality of life still remains a problem even after 80 years. Therefore, the solution to the disease was seen as the ability of individuals with T1D7 to produce insulin again. Previously, pancreas transplantation, insulin-producing beta cell transplantation and pancreatic islet transplantation were tried in sick individuals. Thanks to developing technology and increasing knowledge about stem cells, beta cell differentiation, especially from mesenchymal stem cells (MSCs), and transplantation to individuals with TlD have been tried. In order to increase the differentiation efficiency of stem cells and show better beta cell properties, gene cloning technologies have been used and various genes have been transferred to these cells [3]. 3D cell culture techniques, which emerged later and offer the opportunity to better mimic the body, also influenced TlD studies, and researchers tried to obtain tissues that can produce insulin. Every tissue in the body has a special biocomposition that enables cells to perform their duties perfectly. Although the production of organ matrices with 3D printing technique has become quite a phenomenon in recent years, the biocomposition of the tissue scaffold to be produced is quite different from the natural composition existing in the tissues in the body. Therefore, capturing the content in question in the tissues written with 3D bioprinters, ensuring that the cells cultured in the material used fulfill the correct biological functions, and subsequently converting the produced tissues into clinical application are important issues waiting to be solved. One of the latest innovations has been the cultivation of islet-like structures on decellularized liver extracellular matrix and attempts to create 3D tissues that can secrete insulin [4]. In addition, reshaping the extracellular matrix obtained after decellularization by using it as a raw material in SB-bioprinters is a newly emerged technique, and adipose, cartilage, and heart tissue templates have been tested in vitro [5] and a study on cardiac tissue production only involving transplantation to animals was included. [6]. All of these alternative TlD treatment approaches involve beta-cell replacement, which can reverse the outcome of the disease by replacing destroyed beta cells in the diabetic pancreas. However, it is noteworthy that aggressive autoimmunity still exists after beta cell replacement, and if this is not prevented, specific immune attacks against newly transferred beta cells will continue. Therefore, beta cell replacement alone is insufficient without eliminating existing autoimmunity against beta cells. In order to create Treg-based therapy approaches for autoimmune diseases, induction of Tregs in vivo or in vitro has been carried out in many studies. In these studies, Tregs were obtained as antigen-specific (inonoclonal) or without any antigen specificity (polyclonal). It has been shown that the therapeutic effects of polyclonal Tregs remain weaker than antigen-specific Tregs when transplanted to target autoimmunity [7]; [8]; A promising approach with polyclonal Tregs is the proliferation of the Treg population by low-dose IL-2 administration and in vivo [15]; [16]; [17]. However, the use of polyclonal Tregs propagated in vitro or in vivo carries the potential to induce immune suppression, in other words, the risk of suppressing the beneficial immune system. Since Is will not suppress the entire immune system by providing the advantage of antigen specificity, many studies have taken their place in the literature, including TlD7, in which the application of autoantigen-specific Tregs to mice with autoimmunity for the treatment of autoimmune diseases has been shown to regress the disease. In preventing the onset of the disease, islet antigen-specific Tregs are more effective than polyclonal Tregs. has been shown to be effective. Moreover, it can stop the onset and ongoing diabetes only by the transfer of autoantigen-specific Tregs. Recently, approaches for the production or induction of Tregs in vitro and in vivo have emerged. One of the approaches to in vitro autoantigen-specific Treg production is viral TCR gene transfer to T cells. Stimulating TCR and/or FoxP3 gene-transmitted naïve CD4+ T cells in the presence of TGF-ß can convert these cells into Treg cells. Anti-CD3/anti-CD28-coated beads or antigen-loaded dendritic cells can be used in the presence of high concentration 1L--2 to multiply Tregs in vitro. Limitations in applications have led researchers to in vivo Treg induction. For this purpose, Treg7s can be activated from naïve T cells in the periphery using TGF-β. Although induction via -beta is a frequently used method, it has been determined that the suppression abilities of Tregs obtained in this way are not permanent [26]; [27]. In preclinical studies conducted with animals with TlD, induction of autoantigen-specific Tregs was achieved with antigen-loaded dendritic cells in vivo [28] or in vitro [29]. Antigen-specific tolerance of dendritic cells (antigen-specific tolerance) was achieved through Tregs. Clinical trials have been conducted for the tolerance-based treatment of autoimmune diseases, including TlD [30], and its safety for T1D7 has been tested in a phase I clinical trial [31] by conjugating autoantigens to splenocytes or nanoparticles (creating a chemical bond) and has recently been used for the treatment of autoimmunity and transplantation. A new therapeutic trend has emerged in which immunity can be targeted [28]; [32]; [33]. It was developed in some way and a Sun 1 clinical study on multiple sclerosis was conducted [34]; [35]; [36]. In this technique, poly(lactic-co-glycolide) (PLG) is one of the most used nanoparticles with its biocompatible and biodegradable properties. It is thought that when PLG or splenocytes conjugated with antigens via ECD1 are transferred to living organisms, ECD1 induces apoptosis and therefore the antigen-splenocyte or antigen-PLG therapy mechanism depends on the production of TGF-beta by phagocytes and apoptotic cells [37]; [38]. This approach is thought to prevent autoimmunity. Induction of autoantigen-specific Tregs in multiple sclerosis models and Type 1 diabetes models has been shown to be significantly more effective in its prevention and treatment compared to therapies using only soluble antigens or broad-spectrum immunosuppressive drugs without the use of splenocytes or PLG nanoparticles [32]. It is a highly effective approach with exciting preclinical results [37]. Even if the damaged tissue is restored as a result of targeting Tregs with the above-mentioned effective mechanisms, the autoreactive effector memory T cells present in the treated individuals may show resistance and these remaining cells may cause relapse of the disease. 39]. Therefore, for long-term treatment of autoimmune diseases, the autoreactive immune system in the patient must first be destroyed and then immune tolerance must be achieved. A study and patent of the related study that provide a method for tolerating the immune system of a person exposed to an autoimmune disease have shown that tolerance to autoantigens is restored by first destroying the impaired immune system in animal models of autoimmunity and then by stimulating antigen-specific regulatory T cells. In this regard, in order to destroy the defective immune system, immune cells are subjected to apoptosis with antibodies or low dose gamma irradiation, and then phagocytes are given with disease-related autoantigens for autoantigen-specific Treg induction. This will not be sufficient, because the deficiency in the number of cells that secrete insulin in response to glucose as a result of beta cell destruction also requires a solution. a problem Technical Problems in Current Applications and Technical Solutions Offered within the Scope of the Invention Type 1 diabetes is a disease that causes beta cell deficiency and subsequent irregularities in blood sugar control as a result of the development of an autoimmune response against insulin-producing ß-cells in the pancreas. In terms of TlD treatment, immunotherapy addresses the root cause of TlD by resetting the balance between autoimmunity and regulatory mechanisms. T regulatory cells play an important role in this immune intervention [41]. An alternative treatment for TlD involves beta-cell replacement, which can reverse the outcome of the disease by replacing destroyed beta-cells in the diabetic pancreas. However, it is noteworthy that if aggressive autoimmunity still persists after beta cell replacement, specific immune attacks against newly transferred beta cells will continue. Therefore, beta cell replacement alone is insufficient without eliminating the patient's existing autoimmunity against beta cells. On the other hand, immunotherapy alone will not be sufficient to be applied to individuals with T1D because the deficiency in the number of cells that secrete insulin in response to glucose as a result of beta cell damage will still remain a problem that requires a solution [42]. With immunotherapy (dendritic cells or antigen-conjugated biodegradable nanoparticles- There are already existing approaches to try to eliminate the autoimmunity problem of T1D [43]. However, these studies only target immunotherapy, although many applications have been tried to replace damaged ß-cells in T1D, three of these strategies are preliminary. have come to the fore: (i) cell transplantation, (ii) islet transplantation and (iii) pancreas transplantation [44]. However, pancreas and islet transplantation is both limiting in terms of the source of insulin due to the difficulties in finding donor organs and because it requires lifelong suppression of the immune system. It poses serious problems. Immunosuppressive drugs have been used to overcome the latter condition. However, the use of immunosuppressive drugs has also significantly inhibited the functioning of the immune system, and these people have significant disadvantages in the fight against microbial diseases. Both problems were solved by beta cell differentiation and obtaining insulin-secreting tissues from autologous MSCs taken from patients with TLD. Within the scope of the invention, insulin-producing cells were obtained by obtaining insulin-secreting tissues with stem cells (autologous MSCs) belonging to the patient you used. Within the scope of the invention, insulin-producing cells were obtained. Since the cells are the person's own cells, an immune response does not develop against them, which eliminates the need to suppress the immune system. Type 1 diabetes mellitus is a disease that currently has no cure and has serious effects worldwide. The discovery of insulin has been an important development for diabetic patients. Insulin that cannot be produced sufficiently has begun to be met exogenously. Therefore, TlDM is also called insulin-dependent diabetes mellitus (IDDM). Although TlDM patients can live their lives thanks to the insulin they receive externally through injection, their quality of life is still low. A problem is maintaining its temperature. This problem has been solved by transplanting tissue parts that can secrete insulin according to blood glucose level to sick individuals. After transplantation, patients do not need exogenous insulin. Cell transplantation, islet transplantation and pancreas transplantation contain questions awaiting answers, such as the question of whether or how many of the cells given to the circulation migrate to the desired region, which cell type will be chosen and where the application site will be. However, in the method we use, the tissue scaffold we create not only creates a microenvironment for the cells, but also serves as a vector for them. In this way, cells can be transferred to the desired area of the patient. Production of organ matrices with 3D printing technique has become very popular in recent years. However, the biocomposition of the tissue scaffold to be produced is quite different from the natural composition existing in the tissues in the body. Every tissue in the body has its own biocomposition. This composition has a special design that enables the cells in the tissues to perform their functions perfectly. However, capturing the content in question in textures written with 38 printers poses a big problem. However, using the decellularized pancreatic matrix used within the scope of the invention as an organ writing material in 3D printing preserves the natural biological content of the tissue. This approach, which can play vital roles in ensuring that the cells cultured in the material perform the correct biological functions, also overcomes extremely important problems. Producing glucose-sensitive beta cells that can secrete insulin from stem cells is a very difficult process due to low efficiency. It is also very difficult to obtain human beta cells suitable for transplantation. Although the highest efficiency in transferring genes to cells to gain the desired phenotypic and functional features or to stimulate them in a certain direction can be achieved with viral methods, viral methods are not suitable for clinical use due to safety problems such as the risk of mutagenesis and immunogenicity. On the other hand, non-viral gene transfer vectors create a safer profile in tissue engineering. However, in non-viral methods, the expression of the transferred gene in the target cell could not be reached to the same level. This shows that vector design is very important. An ideal gene transfer vector to be used in tissue engineering should be biocompatible, biodegradable, minimally cytotoxic, and be able to effectively transfer DNA into the cell. It should also be able to maintain the expression of the target protein for the required period of time. The use of gene therapy strategies with a tissue scaffold has led to the emergence of a new field: Gene-activating matrices (Gene-Activated Matrix, GAM). GAM is the direct and continuous delivery of nucleic acids from a tissue scaffold to cells to ensure effective and durable non-viral gene transfer into the cell. In general, the nucleic acids that are desired to be transferred to cells (a plasmid DNA carrying target genes, usually called pDNA) are enclosed in cationic polymers or cationic lipids and mixed by adding them to the solution where tissue scaffolds will be created. Then, SB tissue scaffolds are created from materials that have the ability to transfer genes. The structures are called GAM. When the cells are planted and cultured on GAM, the cationic components loaded into the matrix combine with the cell membranes and thus the genes in the cationic components are transferred to the cells by a non-viral method [45]. Chitosan is thought to be one of the most suitable non-viral methods for this purpose. In particular, the development of the low molecular weight water-soluble oligochitosan molecule has significantly increased the recycling efficiency and has been tested in MSCs [46]. In this invention, the problems related to producing beta cells that can secrete insulin sensitively to the level of glucose from stem cells were solved by obtaining a gene-activating matrix from the natural pancreatic extracellular matrix transferred with chitosan-pDNA nanoparticles and creating a tissue scaffold by writing this matrix on 3D bioprinters. Thus, PdX-1 and MafA [47] genes, which play important roles during pancreatic development to induce ß-cell differentiation, were transferred to MSCs cultured on tissue scaffolds. By stimulating ß-cell differentiation from MSCs with the gene-activating matrices created in this invention, a new generation beta cell differentiation method for TlD has been developed and the technical problems in beta cell differentiation in existing methods have been overcome by providing a GAM-mediated 3D microenvironment produced within the scope of the invention. si One of the most difficult problems has been autoimmunity against insulin-producing cells. To solve this problem, the cells were encapsulated using encapsulation techniques and thus their direct contact with autoreactive T lymphocytes was prevented. However, these methods used did not allow for long-term treatment. Therefore, autoreactivity maintains its hot spot on the agenda after encapsulation. However, with the dendritic cells and antigen-conjugated biodegradable nanopatricles we used within the scope of the invention, specific tolerance to the autoantigen was achieved through Treg programming, and then the biochip containing insulin-producing cells was transplanted. Thus, this problem has been overcome by immunotherapy performed through antigen-specific tolerance of autoreactive T lymphocytes. The low frequency of Treg in peripheral blood has been identified as one of the major challenges to be overcome in Treg-based immunotherapy in the past years. Approaches to induction have emerged. One of the approaches to in vitro autoantigen-specific Treg production is viral TCR gene transfer to T cells. However, these Tregs, which have very high in vivo suppression properties, are not clinically applicable due to transgenic TCR manipulation. In vitro autoantigen-specific Treg production. Difficulties in determining the antigen specificity of Tregs and obtaining a sufficient number of antigen-specific Tregs are other factors that limit clinical applications. situations. These limitations have led researchers to in vivo Treg induction. In this direction, in Viva autoantigen-specific Tregs are generated from naïve T cells in the periphery via TGF-beta [27]; [14] or induced by antigen-loaded dendritic cells [28]. However, the fact that the studies were conducted when autoimmune diseases were not yet developed causes the therapeutic effectiveness of these applications to not be determined. The antigen-PLG or splenocyte therapy approach in the prevention and treatment of autoimmunity has been shown in various animal models to be significantly effective compared to therapies using only soluble antigen or broad-spectrum immunosuppressive drugs without the use of PLG nanoparticles or splenocytes [32]. Inducing autoantigen-specific Tregs in this way is a highly effective approach with exciting preclinical results in multiple sclerosis models and Type 1 diabetes models [37]. On the other hand, in case of disease, immune cells that respond to autoantigen become uncontrolledly activated by increased pro-inflammatory cytokines, causing Tregs to lose their suppression ability. Tregs can even turn into effector cells such as Thl7 cells in this pro-inflammatory environment [24]. Even if the damaged tissue is restored as a result of targeting the cells with the effective mechanisms mentioned above (such as antigen-PLG and dendritic cells), the autoreactive effector/memory T cells present in the treated individuals may resist and these remaining cells may cause relapse of the disease [39]. In order to solve this problem and long-term treatment of autoimmune diseases, the autoreactive immune system in the patient must first be destroyed and then immune tolerance must be achieved. According to the data of the International Diabetes Federation, it is expected that the dollar related to diabetes will increase in 2015. TlD is a disease that results in insufficient insulin production as a result of autoreactive T lymphocytes attacking insulin-producing pancreatic ß cells. Therefore, patients with T1D must take commercially available insulin daily to meet their insulin needs. In addition, chronic TLD disease brings with it very serious secondary complications such as eye damage, neuropathy, nephropathy, heart and blood vessel diseases. When examined from this perspective, diabetic patients face serious economic burdens due to other diseases caused by diabetes. In the document, a method for the preparation of autologous dendritic cells obtained through mesenchymal stem cells is mentioned. During the production of dendritic cells capable of providing immunosuppression, mesenchymal stem cells were used as a stimulator, that is, a supportive stimulus, for dendritic cells to acquire the immunosuppression function. In order for dendritic cells to stimulate mesenchymal stem cells, these two cell types were put into co-culture. In the Chinese patent document numbered CN105670990, one of the known applications in the art, the tissue engineering application and preparation method for directional differentiation developed from mesenchial stem cells is mentioned. The invention was made in the field of biomedical engineering and involves the preparation and application of a tissue engineering material to stimulate the differentiation of mesenchymal stem cells. Poly(lactic-co-glycolic acid), called PLGA, is used, and this polymer is in the class of non-natural polymers, that is, polymers that are not produced in the body and do not exist in the body [49]. Although biocompatible, this polymer is not produced in the body and is a completely foreign environment for cells. As is known, organs consist of two components: Cells and intercellular substances called extracellular matrix [50]. In the decellularization technique used within the patent, cells are removed from the extracellular inatrices and only the natural extracellular matrix of the organ or tissue is obtained. Thus, the polymers used are obtained after organ decellularization and are the organ's own environment that already exists in the body, and they consist of collagen type 1, collagen type 4, fibronectin, glycosaminoglycans, etc., rather than consisting of a single polymer such as PLGA. It consists of natural biomaterial with a complex content containing many natural polymers [51]. Replacing dead beta cells is not a permanent and long-term solution in the treatment of TlD. Because autoimmunity against beta cells still continues in the body. As a result, the permanent solution to T1D9 is not only to complete the beta cell deficiency, but also to eliminate the existing autoimmunity. However, patent number CN105670990 only targets beta cell replacement. It has not been shown that high blood glucose levels are reduced to normal values after the beta cells produced in the patent numbered CN105670990 are transplanted to a diabetic individual, and no m vivo (performed in a living organism) study is included. In short, the function of the product in question has not been demonstrated live. The document mentions a mesenchymal stem cell composition derived from autologous and allogeneic adipose tissue developed for the treatment of diabetic wounds. Diabetic wounds are secondary vascular complications that develop in individuals with diabetes due to excessively increased blood glucose as a result of the disease. It is aimed at wound healing focused on correcting the vascular system (vessel) and inflammation. Autologous mesenchymal stem cells have been used for wound healing and inflammation prevention. It is a known feature of mesenchymal stem cells that they inhibit immune responses in the transplanted area [52]. Mesenchymal stem cells have a high potential for use in clinical treatments today, especially with their ability to be easily obtained from many tissues, their anti-apoptotic and anti-inflammatory properties, their lack of immune response in allogeneic transplantations, and their ability to differentiate into many somatic cells, including ß-cells. Within the scope of the invention, the composition and a biodegradable tissue scaffold are transplanted into the human body. In the Chinese patent document numbered CN104353115, which is one of the known applications in the art, a kit and preparation method for pancreatic decellularized tissue scaffold and the reseed method of the tissue scaffold are mentioned. The tissue scaffold used in the invention is the extracellular matrix obtained from decellularized pancreas. The permanent solution of TlD not only compensates for the beta cell deficiency, but also targets only beta cell replacement, like the middle number of the existing autoimmune unit. It is done via line. However, it is reported in the literature that this cell line is not a pure beta cell line [53]. But more importantly, this cell line is an insulinoma cell line. In other words, it is a cell line obtained from a tumor originating from the B cells of the pancreatic islets of Langerhans. The information in question is also included in the ATCC (American Type Culture Collection) where this cell line is sold [54]. Therefore, no tissue, chip, etc. product produced with these cells is suitable for clinical use today. In the patent numbered CN104353115, just like in the patent numbered CN105670990, the beta cells produced are It has not been shown that high blood glucose levels are reduced to normal values after transplantation to a diabetic individual, and it is not included in any in vivo studies. In short, the function of the product in question has not been demonstrated in vivo, cell stimulation biochip and differentiation of stem cells. In the United States patent document numbered USZOl3017l75, one of the known applications in the art, activated mesenchymal stem cells are mentioned for wound healing and regeneration of damaged tissue. It is a tissue repair method with in Viva application in the initiation of vascularization. The aim is to show how effective the use of mesenchymal stem cells with activated immunosuppression properties can be in preventing inflammation resulting from graft versus host disease (GVHD), which develops as a result of rejection of the transferred tissue (transplant) by the recipient in tissue transplants. In this direction, the in vivo initiation of the activated mesenchymal stem cell composition created to prevent GVHD is the creation of a composition of mammalian mesenchymal stem cells activated with inflammatory cytokines to prevent inflammation and tissue repair, and the transplantation of the cells to this area. This can be any tissue, and the use of the immunomodulatory properties of mesenchymal stem cells in the inflammatory environment is the basic methodology of the invention. It is claimed that its use to prevent islet transplantation may also be functional. In the invention numbered USZOl30l7l75, the cell composition applied in vivo for tissue regeneration and organ repair consists of inflammatory cytokine-activated mammalian mesenchymal stem cells. The application document contains an immunotherapy and kit developed for the treatment or toleration of autoimmune diseases expressed by apoptosis-antigen therapy. The therapeutic steps in the invention are: i) Detection of autoimmune subjects; ii) Directing immune system cells to apoptosis; (a) by administering T cell antibodies (anti-CD3 and anti-CDS) or B cell antibodies (anti-CDZO), (b) or by administering low doses of radiation (radiotherapy); iii) Introducing macrophage cells to the patient following radiotherapy or antibody administration; iv) Finally, the autoantigens that cause autoimmunity are given to the patient. As a result of all these steps, the existing autoimmune cells in the radiotherapy area were eliminated by causing them to undergo apoptosis, and the administered macrophages were enabled to support immune tolerance more strongly when they encountered apoptotic cells. After preparing this environment, an immunotherapy in which Tregs are stimulated by administering autoantigens associated with autoimmune disease to the patient, which has recently been a common method in antigen-specific therapy, has been presented. When both patents are evaluated, the common aspects found are that in the immunotherapy applied for TLD, Tregis are transferred together with macrophages and macrophages specific to the in vivo antigen, whereas in the present invention, antigen-specific dendritic cells are produced by loading (stimulated) onto dendritic cells in vitro, instead of giving autoantigens to patient individuals. and/or conjugated to biodegradable nanoparticles (PLGA-Antigen). By administering the resulting antigen-specific dendritic cells and/or PLGA-Antigen nanoparticles to the patient, the Tregs in the immune system were programmed specifically for the antigens we selected. The purpose of the invention is only immunotherapy; immunotherapy alone will not be sufficient to be applied to individuals with TlD, because the deficiency in the number of cells that secrete insulin in response to glucose as a result of beta cell destruction will still remain a problem that requires a solution. Brief Description of the Invention This invention aims to eliminate autoimmunity in Tl Diabetes patients. It is a TlD treatment method that aims to eliminate it through reprogramming and to provide insulin-producing autologous cell replacement. In line with this purpose, the invention aims to prevent the autoimmune immune system that attacks the insulin-producing ß-cells in T1D, by means of Viva programming of T regulatory cells (Treg). On the other hand, in order for the produced tissue-engineered biochip (insulin-secreting tissue) to continue its function in the in vivo environment, an environment in which autoimmunity can be prevented can be created before tissue transplantation. The invention also includes the production method of an endocrine pancreas-like functional tissue piece that can bring blood sugar to a healthy level by secreting insulin in response to glucose by developing new generation beta cell differentiation techniques in endocrine pancreas engineering. In this respect, the invention includes both a product and the production method of this product. Preparation of the elements that form the insulin-secreting tissue (mesenchymal stem cells, beta cell gene transfer system and decellularized pancreatic matrix: dpESM), printing these elements using 3D bioprinters and creating a tissue. The aim is to obtain the tissue template and culture this tissue template in vitro. Thus, stimulation of differentiation of mesenchymal stem cells in the endocrine-panrectic direction was achieved by the influence of many factors [(i) dpESMa (ii) gene transfer system related to ß-cell differentiation and (iii) endocrine medium culture]. The invention is an endocrine pancreas-like functional tissue fragment product created with a 3D bioprinter, Gene Activating Matrix (GAM; dpESM + cationic polymer-pDNA nanoparticles) consisting of decellularized pancreas integrated with a cationic polymer-pDNA nanoparticulate gene transfer system suitable for differentiating mesenchymal stem cells into beta cells. ) also includes the product. GAM, which stimulates differentiation in this endocrine direction, is a sub-product created within the scope of the invention and is one of the subjects of the invention. On the other hand, in the invention, the biochip (insulin-secreting tissue) produced was transferred to an individual with T1D whose autoimmune attacks were prevented. After all these processes, the ultimate goal of the study is to reduce blood sugar to normal levels thanks to the biochip transplanted to patients with T1D whose autoimmune attacks were prevented. Detailed Description of the Invention A subject of this invention is; It is a therapeutic method for TlD disease. The present invention offers a treatment method that enables ß-cell regeneration in T]Diabetes while also reorganizing autoimmunity, thanks to the 3D culture environment. While ß-cell differentiation from MSCs is carried out on the tissue scaffold produced to provide a microenvironment very close to natural, autoreactive T cells against these cells; It involves the reregulation of dendritic cell- and/or antigen-conjugated biodegradable nanoparticle-mediated Tregs. Therefore, the invention can target the treatment of TlD disease as a whole. Thus, a new strategy is provided that allows the treatment of TlD disease by producing personalized tissues and ensuring the tolerance of autoimmune T cells. The invention Another issue is that it includes a new generation beta cell differentiation method in pancreatic tissue engineering. The tissue scaffold designed in accordance with B-cell differentiation is printed using natural decellularized pancreatic extracellular matrix containing a gene transfer system, and this is the first demonstration of the feasibility of this method thanks to 3D bioprinters. In the tissue produced using pancreas, the natural polymer structure of the matrix (dpESM) is strengthened with non-viral ß-cell gene transfer systems. A favorable 3D microenvironment is provided for beta cell differentiation with the gene activating matrices obtained in this way, and thus a new generation beta cell differentiation method is developed. . The invention, as a tissue engineering application and preparation method, mainly includes the following innovation steps: a) Turning the product obtained by lyophilization of decellularized pancreas (dp-ESM) into a printable material in 3D bioprinters. b) Bioink (Bioink: dpESM + chitosan). -pDNA nanoparticles + MSC composition) Preparation o Integration of the nanoparticulate gene (genes related to ß-cell differentiation) transfer system to dpESM, which will be created by using a non-viral, that is, a cationic polymer instead of the virus. o Obtaining Gene Activating Matrix (GAM), which provides stimulation of stem cells towards the endocrine pancreas, with the combination of chitosan-pDNA nanoparticles ß-cell genes transfer system and dpESM o MSC compound was added at a certain concentration and environmental conditions to the dpESM mixture containing the chitosan-pDNA nanoparticles ß-cell genes transfer system. c) Printing of Insulin-Secreting Tissue (Biochip) from Bioink with a 3D printer. Another subject of this invention is the creation of a treatment product for T1D. Endocrine pancreas-like functional tissue is transplanted to bring blood sugar to a healthy level in individuals with T1D whose autoimmune attacks are prevented. This "product", created by the development of many techniques, constitutes a new concept in itself and the production of endocrine pancreas-like tissue grafts suitable for lowering glucose-sensitive blood sugar in patients with TLD (Type 1 Diabetes). The outlines and content of the method used within the scope of the invention, in which transplantation in the form of subcutaneous biochips is targeted, are as follows: - After the subjects with T1D are identified*, patient-specific selection of TlD autoantigen (ß-cell antigens) combinations and doses to be administered as intravenous injection to subjects with T1D by antibody test. , - Obtaining immature/naïve dendritic cells from bone marrow progenitors or blood mononuclear cells of T1D° subjects in order to ensure antigen-specific tolerance in T1D° subjects, Autologous immature/naïve dendritic cells that will induce tolerance to antigen-specifically program the Tregs existing in the patient in T1D71 individuals. It is obtained from bone marrow progenitors by culture containing GM-CSF cytokine. In short, bone marrow (KI) cells are obtained from femur and tibia sources and erythrocytes are subjected to hypotonic lysis. KI cells are cultured in 10ml medium containing cytokinin. Half of the medium of the cells is renewed every day with fresh medium containing cytokine. In order to determine the purity of the dendritic cells, the cells are marked with CD1 10, a dendritic cell marker, on the 7th day of culture and analyzed by flow cytometry. The obtained dendritic cells are previously The prepared lysate (apoptotic bodies of ß-cells) and GAD65 peptide are co-cultured in the next mentioned step when the Dendritic cells phagocytose the apoptotic bodies and peptides, meeting the term primed/pulsed with targeted antigens - in order to ensure antigen-specific tolerance in subjects with TID. In vitro loading of determined TlD autoantigens into dendritic cells obtained from subjects and/or conjugation to synthetic biodegradable nanoparticles***, Preparation of Lysate (apoptotic bodies of IS-cells), Firstly ready ß-cell line or 3D Biochip with physical homogenization to induce ß-cells to undergo apoptosis. 3<105 cells/well are planted in each well of a 24-well petri dish containing 500 pl of medium. After irradiation with ultraviolet B (UVB, 10mJ/m2) for 45 minutes, it is cultured overnight in a 5% CO2 37°C environment. Characterization of apoptosis is confirmed in Ilow cytometry by annexin V-fluorescein isothiocyanate (FITC) and propidium iodide staining. Autoantigens (ß) of dendritic cells. -cell antigens) co-culture and characterization. 106 cells/well for co-culture of the obtained Dendritic cells with apoptotic bodies and GAD65 peptide as ß-cell specific antigens. Dendritic cells: 3x105 cells/well with apoptotic beta cells (3:1 ratio). It is co-cultured in 24-well plates. SuM synthetic GAD65 peptide is added to the prepared culture and the dendritic cells are loaded (primed) with diabetogenic antigens (beta cell apoptotic bodies and GAD65 peptide) and CFSE is characterized before co-culture. After analysis of Dendritic cells labeled with CD11 after co-culture with, Dendritic cells loaded with apoptotic bodies were determined as CD110- and CFSE positive, while unloaded Dendritic cells were determined as CD11c + and CFSE negative. For the characterization of dendritic cells that will show immune tolerance; After activation by the addition of liposaccharide to the cultures, CD4O and CD86 co-stimulant expression and the release of pro-inflammatory cytokines are analyzed. Dendritic cells that tolerate increased co-stimulant expression based on their resistance to the release of pro-inflammatory cytokines are characterized by flow cytometric analysis and the medium by ELISA. Production of ß-cell Specific Antigen-Conjugated Nanoparticles Biodegradable PLG nanoparticles, which offer the opportunity to conjugate autoantigens and possible donor MHC antigens, ß-cell apototic bodies and ß-cell apototic bodies isolated with the chemical ethylene carbodiimide (ECDI) as ß-cell specific antigens, as mentioned in the previous steps. It is conjugated with the commercially available GAD65 peptide, thus creating a biodegradable antigen-conjugated-nanoparticle (antigen-ECDI-PLG nanoparticle). Briefly, ECDI [(1-Ethyl-3-(3' dimethylaminopropyl)carbodiimide HCl) is used to conjugate ß-cell specific antigens to biodegradable PLG nanoparticles in vitro. PLG particles (3mg) are washed 3 times with PBS to remove sugars from lyotylation and 1 hour with 30mg/ml ECDI and mixed with 1200 tig/ml lysate (apoptotic Bodies of ß cells) and 5 µM GAD65 peptide for each dose. Coupled particles are washed twice with PBS to remove excess ECDF and passed through a cell strainer for 40 µm. The yield is characterized by measuring the amount of protein remaining in the supernatant by protein assay. Flow cytometry, in which PLG nanoparticles are conjugated with CFSE-labeled GAD65 and CFSE-labeled ß cell apoptotic bodies before becoming lysate. Autoimmune disease in T1D patient subjects to achieve antigen-specific tolerance. Inducing system cells to undergo apoptosis with low dose (suplethal level) radiation, - Then injecting dendritic cells and/or antigen-conjugated biodegradable nanoparticles intravenously (IV) to subjects with T1D, using ß-Cell specific antigens (GAD65 peptide and lysate). For loaded dendritic cell application, approximately 2-3 million/injection Macrophage/Dendritic cells can be injected for each mouse, approximately 8-12 million/injection Macrophage/Dendritic cells can be injected for humans, and the dose and number of injections can be increased according to the immune response of the patient. In antigen-conjugated-nanoparticle-mediated tolerance induction, each patient is transplanted with antigen-loaded nanoparticles (nanoparticle/injection containing approximately 10 µg peptide for humans, at least 30 µg in total, 1-2 µg/injection for mice) and the dose and number of injections are determined by the patient. It can be increased according to the immune response. - While the above processing steps are carried out, decellularization of the pancreas is performed and cell-free pancreatic extracellular matrix (dpESM) is obtained. , Germany) is carried out under sterile conditions . The isolated pancreas (dpESM, which does not develop an immune response, can be isolated allogeneically or xenogenically from rodents such as rabbits, rats, mice and other mammals, and the pancreas of human cadavers) was connected to the perfusion system and retrograde (reverse direction) via the hepatic portal vein in order to disrupt the cells and remove them from the organ extracellular matrix. ) perfusion method is used. For this, firstly, 1% TritonX-100 is perfused through the portal vein at 10 ml/min for 60 minutes. Then, 1% is perifused for 120 minutes. Then, again with 1% TritonX-100 at 10 ml/min for 15 minutes. It is perfused for minutes. Sterilization of the matrix is achieved with 300 m1 of 0.1% peracetic acid. Enzymes and chemicals are removed from the tissue by washing the tissue with phosphate buffer (PES) with a pl-Psi of 7.4 at 2 ml/min for 4 hours. For dpESM characterization, firstly in dpESM, Whether DNA content remains is determined by the PCR method. After the decellularization process, histochemical and immunofluorescent analyzes are performed to determine whether the parenchymal and stromal integrity of the pancreas is preserved in dpESM, whether any cells remain, and also the glycosaminoglycan (GAG) content, and it is compared with the natural pancreatic tissue. - Afterwards, the product obtained by lyolyzing and pulverizing the decellularized pancreatic matrix (dpESM) into a printable solution form material in 3D bioprinters, lyophilization and powdering of dpESM; A lyolyzer (FTS Systems Bulk FreezeDryer Model 8-54) is used to completely remove the water retained by dpESM. Decellularized pancreatic tissue, which is kept wrapped in aluminum foil to prevent freezing at -200 C0, and all samples lyotilized in a lyophilizer for 20 hours (±2 hours) between 0 and -100 CJ, are stored in airtight closed packages. It is then sterilized by e-beam (electron-beam) irradiation at 22 kGy. The lyophilized layers are immersed in 0.9% saline for 5 minutes. To pulverize the dpESM, the lyotilized layers are crushed in a pressurized mill with liquid nitrogen. The dpESM powder is prepared by acid extraction of these particles for 3 hours at room temperature. Read more Then, it is washed with sterile distilled water by centrifugation at 1000g at 4°C for 10 minutes. The dried dp-ESM is thoroughly fragmented in liquid nitrogen with the help of a mortar and mixed with 10% pepsin (W/W) prepared with 0.5 M acetic acid for 48 hours, using 10x PBS. The solution is filtered through filters with 40-unit pore diameters to avoid any undissolved matrix particles. The final concentration of the dissolved dpESM is adjusted with 0.5 M acetic acid. Then, mycoplasma screening is performed with commercially available kits (such as MycoAlertTM Myeoplasma Deteetion Kit), the pH of the dpESM solution is expected to be approximately between 2.8-3 at this stage. This pH value is adjusted to 7.4 using 10M NaOH. All these processes are carried out at 10°C to prevent the solution from gelling. It is preserved in liquid form to obtain bioink. Meanwhile, using a cationic polymer (chitosan or other cationic polymers that are not toxic to the subjects) that enables the transfer of genes related to ß-cell differentiation by a non-viral method, and creating the chitosan-pDNA nanoparticulate ß-cell gene transfer system**** by the ionic gelation method, Oligochitosan (MW 7.3kDa; DD 97%) molecule for Chitosan-pDNA nanoparticles to be used as a gene transfer system is commercially available (as available from Novamatrix, FMC Biopolymer, Norway). The ionic gelation formula of chitosan-pDNA nanoparticles is cationic (+ ) is formed through the electrostatic interaction between chitosan and anionic (-). In summary, chitosan is added dropwise to the aqueous solution containing pDNA and Sodium tripolyphosphate (TPP) dissolved in 1%-2% acetic acid and constantly mixed, creating a cross-link between chitosan-pDNA particles. Chitosan-pDNA nanoparticles that precipitate as a result of ionic gelation are kept at room temperature for 30 minutes for stabilization before use. Obtaining Gene Activating Matrix (GAM; dpESM+chitosan pDNA nanoparticles) by combining the gene transfer system with decellularized pancreatic matrix (dpESM) to differentiate mesenchymal stem cells into beta cells in the tissue to be printed on a 3D bioprinter, autologous mesenchymal stem cells from bone marrow or other tissues ( Bone marrow apirates under appropriate conditions for obtaining MSCs and endocrine differentiation, preparing MSCs in number and phenotype for the production of endocrine pancreas-like tissue (biochip), and isolation of bone marrow (KI)-derived mesenchymal stem cells (BCS) from individuals with TlD. It is placed in heparin tubes. In the sterile cabinet, bone marrow aspirates are washed with HBSS buffer (Hans Balanced Salt Solution) by centrifuging twice at 300g. It is taken into 0.8.1% ammonium chloride and kept in +4C for 15 minutes. After centrifugation and washing with PBS, ng/mL bFGF was diluted with DMEM-FIZ (Dulbecco's modified Eagle's medium-low glucose and F12 supplement) culture medium containing 1% glutamax, added to the flask and cultured in an incubator providing an environment of 37 OC71 containing 5% CO2. is done. The medium of the cells is changed every 3 days and when the cells are 70-80% confluent, adhering cells are removed by trypsinization process and passaged at a ratio of 1:3. At the end of the 3rd passage, the cells obtained are characterized. MSCs obtained from the bone marrow tissue of individuals with TlD were isolated thanks to their adhesion properties to the culture vessel, and the morphological characteristics of the adherent cells were examined by contrast-phase microscopy during the study. Flow cytometric analysis (CD73, CD90, CD73, CD90, CD73, CD90, CD73, CD90, CD73, CD90, etc.) were the surface markers of MSCs to determine immunophenotypic features. CD105 positive and CD45 CD34 negativity) and immunocytochemical labeling (Immunohistochemical Labeling (IHK) and Immunofluorescent Labeling (IF)) studies are performed. Gene expressions are determined by RT-PCR. - Preparation of bioink (bioink) by adding chitosan-pDNA nanoparticles and MKHIs to the printable dpESM form - by providing appropriate conditions in which gelation is prevented. After the previously mentioned technique, which is done at 10 ° C to prevent the dpESM solution from gelling, the following elements are added to the dpESM, which is kept in liquid state to obtain the bioink. added: Chitosan-pDNA nanoparticles; As mentioned before, the prepared chitosan-pDNA nanoparticles and the prepared dpESM solution are gently mixed. MKHiler.' To prevent cells and DNA-containing Nanoparticles from being damaged during bioprinting, they are coated with a protective gel that can be degraded immediately after printing and will not disrupt the interaction of cells with the matrix. For this, before printing, gelatin methacryloyl (GelMA) (10% (weight/volume) and 0.5% (weight/volume) 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1 It is prepared by adding cells to the -propanone photoinitiator (PI) solution at a concentration of 5x105 cells/ml. Bone marrow-derived MSCs in the 3rd passage (P3) are removed from the culture container and added to the mixture containing dpESM+chitosan pDNA nanoparticles at a concentration of 6x106 cells/ml. It is added and mixed gently. With the addition of cells, the dpESM+chitosan pDNA nanoparticles+MSC composition is called bioink (bioink). - Printing of the biochip from bioink with a 3D printer is used to print the 3D tissue scaffold designed to differentiate MSCs into pancreatic ß-cells. A high-resolution (~1.25-50 micron), multi-head 3D-bioprinter with adjustable heating and cooling features, an extrusion-based bioprinter, is used with temperature control at 25°C, 25°C to optimize cell viability and printability of the material. The bioprinting process is carried out at °C, 27.5°C and 30°C and the results are evaluated and the optimum conditions for each gel are determined and printed. The tissue scaffolds created following the bioprinting process are placed approximately 8.5 cm into the printed tissue product to ensure rapid cross-linking. By applying UV light at 850 mW for 17 seconds from a distance, it is cross-linked and gelled (photo-polymerization by photocuring process). In case the cross-linking process cannot be achieved to the desired degree, Alginate equal to the amount of GelMA added to the bioink is added (V/V) and left in 10µM CaC12 solution for 3 minutes after printing. Cell proliferation/viability analysis of the biochip obtained from bioink with 3D-printing Determination of cell viability in the biochip Fluorescent Live/Dead Staining (as in the Live/Dead Cell Double Staining Kit/Sigma-Aldrich commercial product) and [2-(4-lod0phenyl)-3- (4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium] is determined by two separate analyzes with 3D cell culture viability analysis kits (3D Cell Culture HTS Cell Viability Complete Assay Kit, as in the Biovision commercial product). - Obtaining insulin-secreting beta-like cells from MSCs in the tissue by culturing the tissue printed from bioink on the SB bioprinter in endocrine differentiation medium, and differentiation of MSCs into ß-cells inside the biochip; It is both physically induced thanks to the tissue scaffold obtained from the pancreatic extracellular matrix, induced at the gene level thanks to the MafA and Pdx-1 transcription factors carried by the gene activating matrix in the biochip design, and induced by adding inducer factors to the medium. Thus, ß-cell differentiation of MSCs is supported more strongly. For pancreatic ß-cell differentiation, MSCs are induced with a four-step protocol. Differentiation is first performed with 10% Human AB Serum (if the patient is a human, Fetal for other mammalian patient individuals). calf serum is used), cells are then initiated by culturing for 3 days in LG-DMEM medium supplemented with 10 mM nicotinamide and 4 nM activin A, 25 ng/ml recombinant EGF and 0.5 mM ß-mercaptoethanol. In the third induction step, cells are incubated in LG-DMEM medium containing 2% Human AB Serum, 10 mM nicotinamide, 10 HM Exendin-4, 10 µg/ml INGAP-pp and cells, 10 mM nicotinamide, 10 HM Exendin-4. Differentiation is completed by culturing with H-DMEM containing 10 µg/ml INGAP-pp and 1)( ITS and recombinant bFGF. Cells are incubated under controlled culture conditions at 37°C, 5% CO. The media of the biochips are changed and refreshed every two days. MKH" In order to evaluate the differentiation of cells into ß cells at the level of gene expression, Gata-4, HnBb, Hnf4a, insulin I, insulin II, islet amyloid polypeptide (IAPP), glucose transporter-2 (Glut 2), C-peptide, PdX-1, Nkx2. 2, MafA, Ngn3, MafA and Isl-1 genes are evaluated in Real Time- PCR. INSULIN and C-PEPTIDE ELISA analysis (Human C-Peptide ELlSA Enzyme-Millipore) is performed to determine the secreted insulin level, and to see at the protein level, GATA-4, HNF3B, HNF4A, INSULIN I, INSULIN C-PEPTID, PDX-l, NKX2 are analyzed. .2, MAFA, NGN3, MAFA and ISL-1 are determined by immunofluorescent labeling. To characterize the similarity of the biochip scaffold to the parenchymal and stromal structure of the natural pancreas, Hematoxylin-Eosin (H-E), Alcian Blue and Sirius Red stainings are examined by histochemical method. To determine the apoptotic damage status of the cells in the biochip, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) labeling is performed using apoptosis detection kits (such as those from ln-Situ Cell Death Detection Kit, F. Hoffmann-La Roche). For the morphological analysis of the biochip, To determine its function through in vitro glucose-sensitive insulin secretion analysis; the insulin level in the medium of the biochip is determined by a commercial kit (Millipore method). To determine the glucose-dependent insulin release, the ß-cells in the biochip secrete/secret insulin into the medium in vitro, depending on the glucose added to the medium. To test whether they do not secrete, they are exposed to two different glucose concentrations and insulin is determined in the collected medium. Before starting the insulin secretion analysis, differentiated ß-cells are cultured in insulin-free medium for 48 hours, and the wells are washed periodically with PBS until the insulin is completely removed. 5 Serum-free LG-DMEM containing BSA (low glucose content; 5.5 mmol/L) is added and the cells are incubated for 1 hour at 37°C. The supernatant is collected and frozen at -70°C for analysis of basal insulin secretion. Then, the cells are cultured in H-DMEM (high glucose content; 25 mmol/L) medium for 1 hour. At the end of the period, supernatants are collected and frozen to determine glucose-stimulated insulin release. The amount of glucose-stimulated insulin and protein levels in the secreted supernatants of MSC differentiated ß-cells (experimental group), undifferentiated MSCs (negative control) and commercially available ß-cell line (positive control) are determined (in a microplate reader). - It consists of the steps of transplanting pancreas-like functional tissue grafts in the form of subcutaneous biochips to subjects with T1D, and bringing the blood sugar to a healthy level by secreting insulin in response to glucose in the subject with T1D, thanks to the biochip. Contamination Analysis of the Biochip with Bioburden Before Transplantation, and determining the bacterial contamination in the biochip after differentiation. To determine whether it is present or not, 5 ml of the differentiation medium is taken in a 15 ml conical bottom tube and mixed with 7 ml of SOC medium, and its absorbance is determined at 600 n at 37°C. Following immunotherapy, biochips prepared in vitro are transplanted under the skin of TlD individuals under anesthesia. Evaluation of the therapeutic effect of the transferred biochip. In individuals who have been followed for 23 weeks after the transplant, the fasting blood glucose level (fasting blood sugar, ACS) is measured with periodic blood samples. ACS 2 126 mg/dl is considered diabetic values. Elisa analyzes of C-peptide and insulin. On the and 140th days, a serum sample is taken and human-specific C-peptide level and human non-specific insulin level are measured with an ELISA assay kit (Wako Osaka, Japan and Shibayagi, Gunma, Japan). Oral glucose tolerance test The function of the biochip we will transfer is measured on the 30th day with OGTT (Oral Glucose Tolerance Test). How this test was performed: The individual with TLD was fasted for 8-12 hours and given glucose solution orally (adults: 75g glucose-containing solution, children: 1.75g/kg body weight) and at different times (0, 30, 60 and 120 minutes). Blood glucose measurements in diabetic individuals are considered to be 2 126 mg/dl and postprandial blood glucose 200 mg/dl. For this purpose, glycosylated hemoglobin test HbAlc is measured in blood samples taken from individuals after 72 hours with a portable glucometer device (Medisense Precision QlD; Abbott Laboratories, Bedford, MA, USA). (Glycosylated Hemoglobin) test is valuable in the follow-up of diabetic patients and gives an idea about the three-month blood sugar average. The ideal level for HbAle in diabetic patients is considered to be < 6.5%. As a result, a technique has been developed in which autoimmune attacks in TLD can be prevented as a result of tolerance-based immunotherapy, and blood glucose levels can be brought to normal levels and long-term normal blood glucose levels can be maintained thanks to the transplantation of glucose-sensitive insulin-secreting tissue grafts produced. Transplanted tissue grafts prevent autoimmune attacks. The most important feature of this technique in its therapeutic use is its ability to inhibit blood sugar in glucose-sensitive, diabetic individuals. Diagnosis of which type of diabetes antigens the patients have a reactive immune system is also carried out by performing an antibody test. The information obtained here is used for the selection of personalized autoantigens. ** Instead of or in addition to GAD65, which is used as an autoantigen in the method of the invention that aims at personalized treatment. The antigen(s) detected at the highest level with autoantibodies and identified in the development of diabetes can be used as biomarkers of ß cell damage in individuals. For example; non-specific islet cell antigens (ICAs=lslet Cell Antibodies) such as insulin, insulinoma antigen-2 (lA-2) and transmembrane protein tyrosine phosphatase (lCA512), islet-specific glucose-e-phosphatase catalytic subunit related protein (IGRP) (Wenzlau and Hutton, 2013). Which antigen will be selected will be determined by subjecting the individual with TLD to the antibody test used for the diagnosis of T] Diabetes, in the presence of a doctor or veterinarian who has standard experience with this technique. In vitro loading of autoantigens into dendritic cells obtained from subjects and/or conjugating them to synthetic biodegradable nanoparticles) can also be used as an alternative. However, in cases where it is insufficient to prevent autoimmune attacks, they can be used together to increase the immune tolerance effect. In alternative use, the choice of which method to use can be determined by a doctor or veterinarian with standard experience in this field, depending on the ability to obtain dendritic cells from patient subjects. To ensure the immune tolerance of autoreactive T cells causing TlD against ß-cell antigens, tolerant antigen-conjugated nanoparticles are obtained by in vitro conjugation of biocompatible and biodegradable nanoparticles with TlD autoantigens such as polymeric nanoparticles (such as co-glycolide) or liposomal nanoparticles. Following the injection of antigen-conjugated nanoparticles into subjects with TlD, the existing Tregs in the patient are reprogrammed towards tolerance. It is named and pDNA contains Pdx-1 and MafA genes or [3 transcription factors involved in cell development. As a result of combining Chitosan and pDNAs by ionic gelation, nano-sized Chitosan-pDNA particles are obtained as gene transfer vectors. The term "individual" or "subject" mentioned as a subject or individual with TlD refers to any human or non-human mammal. Dosage regimens in the immunotherapy method administered to individuals with TlD* by intravenous injection 24 hours after radiation irradiation determine the targeted optimum response ( e.g., tolerance and/or therapeutic effect established in a subject). For example, a single dose expressed as an injection may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased according to the requirements of the disease state. TlD autoantigens (ß-cell antigens); Ready-made synthetic antigens defined as autoantigens in TlD (GAD65 and/or 3B are antigens obtained from the lysate of insulin-secreting tissue/apoptotic bodies of ß-cells. In vivo antigen-specific programming of Tregs, i) Detection of autoimmune subjects; ii) Apoptosis of immune system cells by applying low doses of radiation (radiotherapy); iii) In vitro loading of determined TlD autoantigens into dendritic cells obtained from subjects and/or conjugating them to synthetic biodegradable nanoparticles in order to ensure antigen-specific tolerance in subjects with T1D°, iv) Following radiotherapy, the application of autoantigen-loaded dendritic cells and/or antigen-conjugated biodegradable nanoparticles to T1D° Giving li balance; The method is carried out in steps. An endocrine pancreas-like tissue piece (biochip) created with a 3D bioprinter obtained by the method of the invention, pancreatic extracellular matrix obtained by lyophilization of decellularized pancreas in its structure, chitosan-pDNA nanoparticles containing genes (PdX-1 and MafA genes or [3] involved in cell development. Other transcription factors involved) include the transport system and autologous mesenchymal stem cells (derived from bone marrow or other tissues). The product created with a 3D bioprinter contains Gene Activating Matrix (GAM; dpESM+chitosan pDNA nanoparticles), which consists of decellularized pancreas integrated with a chitosan-pDNA nanoparticle gene transfer system suitable for differentiating mesenchymal stem cells into beta cells. GAM, which stimulates differentiation in this endocrine direction, is a sub-product created within the scope of the invention and is one of the subjects of the invention. A cationic polymer that enables the transfer of genes related to ß-cell differentiation via a non-viral method; A chitosan-pDNA nanoparticulate ß-cell gene delivery system is created by using chitosan or other cationic polymers that are non-toxic to the subjects. The method of the invention is used therapeutically in the treatment of TlD patients using the said endocrine pancreas-like tissue piece (biochip). The product is transferred under the skin and lowers blood sugar by secreting glucose-sensitive insulin. The product obtained within the scope of the invention is suitable for clinical use since a non-viral cysteine is used for gene transfer. In addition, after the matrix obtained by decellularization of the organ is pulverized, the product can be designed with software in electronic media and produced according to the desired micro patterns with 3D bioprinters, thus it is suitable for integrating the non-viral gene transfer system into the matrix. The product is a matrix that allows the addition of various polymers, growth factors and media stabilizers to the solution obtained by solubilizing the decellularized pancreatic matrix, if needed. The method of the invention allows biochips to be given their 3D final shape in accordance with the redesign of the matrix in the computer environment and the matrix to be designed according to the target region to be transferred when transferred to a living organism. If the cell viability is low in the sample tissue printed from the bioink used as a cellular matrix solution, the solution obtained by mixing the gene transfer system created with the matrix in solution can be used as bioink and printing on a 3D bioprinter can be carried out with these two elements. MKHWer can be added to the scaffold after printing during culture. A doctor or veterinarian with standard experience with this technique can easily determine the effective dose of the components (dendritic cells, antigen-conjugated nanoparticles or antigens) required in immunotherapy. For example, to achieve the desired therapeutic effect through the immunotherapeutic components used, the doctor or veterinarian first administers it at a level below the required level and increases the dosage over time until the desired effect is achieved. In the new generation insulin synthesizing tissue production method, which is one of the subjects of the invention, two separate areas of tissue engineering, namely decellularization and ? By using a combination of bioprinting techniques, a hybrid suppression technique has been developed, producing a bioactive matrix that rewrites the pancreas' own extracellular matrix using stem cells. By differentiating autologous mesenchymal stem cells, insulin-producing beta cells are obtained and the production of an endocrine pancreas-like functional tissue piece that secretes insulin is achieved. Another subject of the invention is the therapeutic method for the treatment of T1D7, by first suppressing the autoimmune immune system that attacks the insulin-producing ß-cells in T1D. After autoimmune attacks are prevented in individuals with TlD, the insulin-producing tissue piece created within the scope of the invention is transplanted to individuals with TlD in the form of subcutaneous biochips. Thus, the tissue can bring blood sugar to a healthy level by secreting insulin in response to glucose in diabetic individuals. Within the scope of the invention, first of all, patients with TlD The patient subjects are identified and the autoantigens and doses to be administered to the subjects with detected TlD are determined to induce antigen-specific tolerance against TlD in subjects with TlD and/or antigen-conjugated biodegradable nanoparticles. Both methods can be used as alternatives. However, it can also be used together to increase the immunotolerance effect when preventing autoimmune attacks is insufficient. In alternative use, the choice of which method to use can be determined by a doctor or veterinarian with standard experience in this field, depending on the ability to obtain a sufficient number of dendritic cells from patient subjects. After the existing autoimmune system cells in patient subjects with TlD are subjected to apoptosis with low dose (suplethal level) radiation, these dendritic cells and/or antigen-conjugated biodegradable nanoparticles are injected into the patient subject with TlD, resulting in TlD in the subject. Induction/reprogramming of T-regulatory cells (Treg) that will provide specific tolerance to the opposing antigen is carried out. On the other hand, for the production of insulin-secreting tissue, which constitutes another part of the invention, the matrix that will form the scaffold of this tissue: cell-purified pancreatic extracellular matrix (dpESM) is obtained by pancreatic decellularization and its lyophilization is carried out. After the water in its structure is completely removed by the lyophilization process, dpESM is turned into powder and then turned into a solution at the appropriate concentration to best support cell viability and to enable bioprinting in 3D printers under the most ideal conditions. Meanwhile, as the second step in obtaining insulin-secreting tissue, a matrix is created that transfers ß-cell genes, which enables the tissue scaffold to also transfer genes, in order to achieve a stronger differentiation from mesenchymal stem cells (MSCs), which are used as cellular elements in the tissue, to insulin-secreting beta cells. . Chitosan-pDNA nanoparticles used as ß-cell gene delivery system; It is prepared by combining chitosan, a cationic polymer, with the pDNA vector carrying ß-cell genes. Mesenchymal stem cells (MSCs), which are the last element of the endocrine pancreas-like tissue desired to be produced, are also obtained at this time and made ready in terms of number & phenotype for endocrine differentiation. To prevent cells from being damaged during bioprinting, they are coated with a protective material that can be degraded immediately after printing and will not disrupt the interaction of the cells with the matrix. To obtain the bioink, chitosan-pDNA nanoparticles and MSCs are added to dpESM, which is preserved in liquid form, and a pre-designed 3D tissue is printed using a 3D bioprinter. If the cell viability is low in the sample tissue printed from the bioink used as a cellular matrix solution, the generated gene transfer The solution obtained by mixing the system with the matrix in solution is used as bioink, and printing is carried out with these two elements in the 3D bioprinter, and MKH71er is added to the post-printing tissue scaffold during culture. In both cases, the tissue scaffold created after printing is Gene Activating Matrix (GAM: dpESM + chitosan- By culturing the 3D tissue obtained in both ways in endocrine differentiation medium, the mesenchymal stem cells in the tissue are differentiated in the endocrine direction and insulin-secreting beta-like cells are obtained. In this stem cell differentiation, a versatile endocrine system is obtained thanks to GAM and endocrine culture medium. differentiation stimulation is provided. After the transplantation of endocrine pancreas-like functional tissue grafts (biochips) in the form of subcutaneous biochips to patient subjects with TLD, it is tested whether the biochip can reduce blood sugar to a healthy level, which will secrete insulin in response to glucose. Subject or individual with T1D; the terms "subject or individual" mentioned refer to any human or non-human mammal that develops TlD. In the invention, the element for which a product for the treatment of which is created and at the same time a therapeutic method is presented are subjects with TlD. TlD 0t0 antigens (ß- cell antigens); These are antigens determined to be loaded into dendritic cells and/or conjugated to biodegradable nanoparticles under in vitro conditions in order to provide antigen-specific tolerance. These are: Ready-made, synthetically produced antigens (such as GAD65) identified as autoantigens in T1D7 from ß-cell antigens, produced within the scope of the invention. They are antigens obtained from the lysate of the 3D tissue and the apoptotic bodies of ß-cells. Antigen-specific tolerant dendritic cells are obtained by in vitro loading of dendritic cells obtained from the blood of subjects with TlD with TlD autoantigens. Antigen-loaded dendritic cells are created to reprogram cells towards tolerance. Dendritic cells are co-cultured with beta cell antigens that cause autoimmunity and are designed specifically for the antigens that cause T'lD autoimmunity, thus ensuring immunosuppression specific to these antigens only. Within the scope of the invention, T regulatory cells (Treg) are stimulated by administering antigen-specific designed dendritic cells to viv07. Thus, to prevent autoimmunity in TlD, antigen-specific immunosuppression is performed by in vivo programming of T regulatory cells (Treg) in an antigen-specific manner. Dendritic cells are transferred to the patient after they are loaded with autoantigens in vitro and become antigen-specific. In other words, TlD Before performing dendritic cell transfer to the subject, dendritic cells loaded (stimulated) with autoantigens are created in vitro. Stimulated tolerant dendritic cells specific to this antigen are administered to patients within the scope of the invention, with biodegradable nanoparticles (PLGA-Antigen) to which the antigen is conjugated, together with the dendritic cell. PLGA-Antigen nanoparticles are prepared with selected antigens (beta-cell antigens) related to TLD. By administering PLGA-Antigen particles to the patient after radiotherapy, Tregs in the immune system are programmed specific to the antigens we have chosen, thus providing antigen-specific tolerance-based immunotherapy. . The antigens used in the invention (e.g. GAD65) are used as antigens obtained from beta-cell apoptotic bodies (in combination when necessary according to the patient). Antigen-conjugated biodegradable nanoparticles; To ensure the immune tolerance of autoreactive T cells causing TlD against ß-cell antigens, biocompatible and biodegradable nanoparticles [polymeric nanoparticles or liposomal nanoparticles such as PLG poly(lactic-co-glycolide), also known as PLGA poly(lactic-co-glycolic acid)]] Tolerant antigen-conjugated nanoparticles are obtained by in vitro conjugation with TlD autoantigens. Antigen-conjugated nanoparticles are created to reprogram the existing Tregs in the direction of tolerance after injection into T1D subjects. Low-dose radiotherapy; existing autoimmune system cells in T1D subjects are subjected to apoptosis with low-dose (suplethal level) radiation for 1 second. Radiotherapy is applied both to eliminate existing autoimmune cells by causing them to undergo apoptosis and to support immune tolerance more strongly when antigen-loaded dendritic cells and nanoparticles administered after radiotherapy encounter apoptotic cells. by loaded dendritic cell and antigen-conjugated biodegradable nanoparticles-mediated Treg induction); Antigen-loaded dendritic cells and antigen-conjugated biodegradable nanoparticles are injected into T1D subjects at certain doses and concentrations to ensure immune tolerance of autoreactive T cells, which are the cause of the disease, against ß-cell antigens. Like this; i) In vivo programming of T regulatory cells (Treg), which play a major role in suppression, to ensure immunotolerance of autoimmune immune cells that attack insulin-producing ß-cells in individuals with T1D and ultimately prevent autoimmune attacks, and ii) Integration of the tissue engineering product (biochip) we created, which is the basis of the invention. Decellularized pancreatic extracellular mantrix (dpESM) enables the creation of an environment in which autoimmunity can be prevented before biochip transplantation in order to maintain its function in the vivo environment; cell-free pancreatic extracellular matrix (dpESM) is obtained by pancreatic decellularization to be used as bioink (material to be loaded into the cartridge) for 3D bioprinting. ) is done Since decellularized material is used in the tissue scaffolds produced with the technique within the scope of the invention, in addition to providing the opportunity for bioprinting with the natural extracellular matrix of the organ, it is also very important in the differentiation of mesenchymal stem cells, which are a member of the pancreatic tissue, into beta cells. Extracellular matrix components are generally conserved structures across species, and even in xenogenic transplants they either do not create any immune response or the immune response they create can be easily tolerated. In order to make the matrix obtained after the pancreas decellularized printable on BB printers; The matrix was lyolyzed and turned into powder and then converted into soluble form, giving it a gelling feature. This produced form of the matrix is very suitable for supporting cell viability and creating a designable microenvironment. The polymers used within the scope of the invention are obtained after organ decellularization and are a medium belonging to the organ that already exists in the body, and are much more than consisting of a single polymer like ready-to-use synthetic polymers; they consist of collagen type, collagen type 4, ibronectin, glycosaminoglycans, etc. It consists of natural biomaterial with a complex content containing many natural polymers [31]. Chitosan-pDNA nanoparticles ß-cell genes delivery system; It is created to ensure the differentiation of mesenchymal stem cells in the produced pancreatic tissue into insulin-secreting ß-cell-like cells. This is a non-viral, that is, nanoparticulate gene (genes related to ß-cell differentiation) delivery system that will be created by using a cationic polymer (chitosan or other cationic polymers that are not toxic to the subjects) instead of the virus. Chitosan-pDNA nanoparticles "pDNA" in the ß-cell gene transfer system is the short name for plasmid DNA containing beta cell differentiation genes, and "pDNA" contains Pdx-1 and MafA genes or [3] transcription factors involved in cell development. Chitosan and pDNA' It consists of nano-sized Chitosan-Gene Activating Matrix (GAM) and dpESM+chitosan-pDNA nanoparticles as a gene transfer vector as a result of combining the proteins with ionic gelation. In order to obtain Gene Activating Matrix (GAM), which provides stimulation of stem cells towards the endocrine pancreas, chitosan-pDNA nanoparticles and ß-cell gene transfer system are integrated with the printable dpESM form in 3D bioprinters. At the same time, the preparation of bioink was continued by combining two elements. With the GAM technology prepared with this technique, beta-cell differentiation from mesenchymal stem cells is strongly supported by both the gene transfer system and the effect of natural dpESM. Mesenchymal stem cells (MSCs) were used as a stem cell to create insulin-secreting cells in the pancreas, in other words Mesenchymal stem cells have been differentiated into beta cells through a series of methods. It is also the third element of the bioink. Within the scope of the invention, autologous mesenchymal cells (taken from the patient himself, from the same individual) are obtained from the patient's bone marrow or other tissues and are directed to be differentiated/transformed into ß-cells. Although the properties of MKH71s such as anti-apoptotic, anti-inflammatory and not causing an immune response in allogeneic transplantations are the reasons why these cells are selected, immunosuppression properties are not the main purpose. . In the invention, the 3D tissue scaffold (Bioink; dpESM + chitosan pDNA nanoparticles + MSC composition) created with autologous and bone marrow-derived mesenchymal stem cells and its creation method are presented as innovations. Bioink (Bioink; dpESM+chitosan pDNA nanoparticles+MKH composition); The elements used as bioink, that is, printing material (material to be loaded into the cartridge) for the 3D bioprinter, are integrated in a printable form in the bioprinter and the preparation of the bioink is completed. Bioink and ? 3D tissue scaffolds are produced using bioprinters. These 33 structures create a structural environment for mesenchymal stem cells, with pre-designed micro patterns and tissue-specific mechanical properties according to natural tissue, to transform into an insulin-secreting pancreatic tissue, and provide biocompatible niches with a porous structure. The cell's response to the material provides important factors such as its adhesion to the material, proliferation on the material, differentiation, and protein synthesis profile. Endocrine pancreas-like functional tissue piece (biochip); It is both a new product created within the scope of the invention and a product used in the T1D treatment method, which is another scope of the invention. In this context, it is a biochip that is transplanted subcutaneously to enable the subject with TlD to return blood sugar to a healthy level by secreting insulin in response to glucose after immunotherapy. The invention enables the reprogramming of the immune system that causes autoimmunity in Type 1 diabetes (TlD) and then reduces blood sugar to a healthy level by secreting insulin in response to glucose. It is a therapeutic method for the treatment of TlD as the production and transplantation of endocrine pancreas-like functional tissue fragments that can increase blood pressure, and it is also the production of natural tissues that secrete glucose-sensitive insulin through the development of new generation beta cell differentiation techniques in endocrine pancreas engineering. The invention consists of two main substudies in detail: immunotherapy and B-cell replacement (replacement of lost tissue). Within the scope of the invention, the entire approach to the treatment of TLD is presented, both as immunotherapy and beta-cell replacement. Immunoregulation approaches are rearranged within the scope of the invention and two important points are targeted: l) T regulatory cells (Treg), which play a major role in immunosuppression, to ensure immune tolerance of autoimmune immune cells (especially autoreactive cytotoxic T cells) that attack insulin-producing ß-cells in individuals with T1D. In vivo programming and ultimately preventing autoimmune attacks. 2) Creating an environment in which autoimmunity can be prevented before biochip transplantation, so that the tissue-engineered biochip (insulin-secreting tissue) can continue to function in vivo, and for beta cell replacement, by developing new generation beta cell differentiation techniques in endocrine pancreas engineering. It provides the production and transplantation of an endocrine pancreas-like functional tissue piece that can bring blood sugar to a healthy level by secreting insulin in response. The steps that provide methodological innovations in the invention are enriched with gene transfer for ß-cell replasmin in T1D, and the tissue in which more than one previously tested technique has been developed. It is about creating engineering products. Here, a hybrid printing technique is developed by using decellularization and 3D bioprinting techniques, which are two separate fields of tissue engineering, and the production of a bioactive matrix that rewrites the pancreas' own extracellular matrix using stem cells is achieved. In light of all this information, this invention includes the following innovations: i. The complete therapeutic approach of the invention for TlD, ii. Production of endocrine pancreas-like functional tissue fragment iii. Tissue engineering method used iv. Transplantation of this tissue: In the present invention, autoimmune system cells in individuals with TlD were subjected to apoptosis with low-dose radiotherapy, and then, by induction of dendritic cells and antigen-conjugated biodegradable nanoparticle-mediated regulatory T cells (Treg), the tolerance of autoreactive T cells in T1D7 (immunity against ß-cells) was achieved. The problems related to insulin requirement in studies targeting only autoimmunity are overcome in this invention as follows: A biochip that secretes insulin sensitively to the glucose produced is transplanted to individuals with T1D in order to compensate for the insulin deficiency caused by ß-cells lost with the development of TlD following the reprogramming of autoimmunity. As a result, a technique is being developed in which the autoinity present in T1D is eliminated, the blood glucose level is brought to normal levels, and long-term normal blood glucose levels can be maintained. The endocrine pancreas-like tissue scaffold production technique, which will be used for beta-cell differentiation from MSCs, is used for the treatment of TlD. It is a specially designed technique. This technique, which allows the solution produced by solubilizing the extracellular matrix obtained after pancreatic decellularization to be printed according to previously designed micro patterns on 3D bioprinters, is a method used for the treatment of TLD for the first time. Thus, by combining the strengths of both the decellularization technique and the 3D bioprinting technique, a new and powerful hybrid technique for endocrine pancreatic tissue engineering is created. Stimulation of differentiation towards the endocrine pancreas of mesenchymal stem cells in 3D tissue, printed from bioink on 3D bioprinters and cultured in endocrine differentiation medium; i.ß-gene transfer system related to cell differentiation, ii. dpESM, and iii. It is provided by endocrine medium. In the invention, cell-free pancreatic extracellular matrix (Decellularized pancreatic extracellular matrix (dpESM)) is obtained by pancreatic decellularization to be used as bioink, that is, printing material (material to be loaded into the cartridge) for the 3D bioprinter. Since decellularized material is used in the tissue scaffolds produced with this technique, it not only offers the opportunity for bioprinting with the natural extracellular matrix of the organ, but also is very important in the differentiation of mesenchymal stem cells, which are a member of the tissue cultured and created in this tissue scaffold, into beta-cells. Extracellular matrix components are generally conserved structures across species and produce either no immune response or can be easily tolerated, even in xenogenic transplants. After obtaining dpESM, the product, which was pulverized after lyophilization, was turned into solution for printing in order to obtain a bioink form that optimizes cell viability and printability of the material in 313 bioprinters. Thus, a homogeneous composition in the produced tissue was enabled while obtaining a form in which the matrix could be printed. After immunotherapy, an endocrine pancreas-like functional tissue piece (biochip) is obtained, which is transplanted under the skin so that the subject with T`lD can bring the blood sugar to a healthy level by secreting insulin in response to glucose. Biochip is a biocompatible and living system, plastic or metal-like. They are living and essentially biological systems that do not contain materials and are designed to be transferred to the living body. Thanks to the biochip produced within the scope of the invention, the aim of the invention is to provide beta cell replacement and to eliminate the autoimmune response by reorganizing the dendritic cell and T regulatory cell interactions. It does not approach the disease from a single point, but sees the disease as a whole and addresses it in a complex and multi-step manner. The tissue scaffold used within the scope of the invention is produced according to the desired micro-patterns with BB bioprinters after the matrix obtained by decellularization of the organ (the similarity of the mentioned patent and our patent in terms of matrix is only up to this point) is pulverized and designed in electronic environments through software, thus making it non-viral. It is produced as a result of an integrated method in which the gene transfer system can also be integrated into the matrix. Using this system brings many advantages; I. The solution obtained by solubilizing the decellularized pancreatic matrix used constitutes the basic material of the tissue scaffold. ii. Thanks to the redesign of the matrix in the computer environment, the final shape of the chips to be produced allows the matrix to be designed according to the region to be transferred when transferring it to a living organism. iii. If the matrix content is needed, various polymers, growth factors, media stabilizers, etc. can be added to the solution obtained by solubilizing the decellularized pancreatic matrix. Allows the addition of . In other words, the system used is a system suitable for enriching the environment. iv. Integrating the gene transfer system, which will facilitate the transformation of the transplanted cells into insulin-producing beta cells after the recellularization of the matrix, into the decellularized pancreatic extracellular matrix solution is also possible thanks to the method used within the scope of the invention. Here, the gene transfer system is preferred as a non-viral system in order to produce clinically suitable biochips. Briefly, the advantages and innovations provided by the invention: In many studies on F TlD, allogeneic beta cell, islet or pancreas transplantation is tried, and since such transplantations are allogeneic, they cause very serious immune responses. Patients have to use immune suppressing drugs. Within the scope of the invention, autologous mesenchymal stem cells taken from sick people are used and beta cell differentiation is performed from these cells. Therefore, the tissues produced are personalized tissues and do not cause an immune response when transplanted into the patient. Individuals with TLD have to use exogenously taken insulin throughout their lives. Thanks to the tissue produced, the body's own insulin need will be produced according to the blood glucose level and sick individuals will be able to continue their lives without the need to use exogenous insulin. Considering that it is quite high, it creates a significant burden for our country's economy. Thanks to the tissue produced, this economic loss is also prevented. Unlike 2D culture methods, beta cells created from BF MSCs are produced in SB environments. This provides the opportunity to better mimic the body, unlike many previous studies. I› The invention will be used for beta cell differentiation from MSCs. The insulin-secreting endocrine pancreas-like tissue scaffold production technique is a technique specially designed for the treatment of TlD. This technique, which allows the solution produced by solubilizing the extracellular matrix obtained after decellularization of the pancreas, to be printed according to previously designed micropatterns on 3D bioprinters, is a technique for the first time in TlD. It is a method used for treatment. Thus, by combining the strengths of both the decellularization technique and the 3D bioprinting technique, a new and powerful hybrid technique for endocrine pancreatic tissue engineering is created. Gene-activating matrices (GASM: dpESM+ chitosan-pDNA nanoparticles) are obtained by strengthening the natural (non-synthetic) polymer structure of the pancreas-derived matrix (dpESM) written on 3D bioprinters and non-viral gene transfer systems (chitosan pDNA nanoparticles). In the tissue scaffold printed from this matrix, a unique microenvironment is provided for beta cell differentiation from mesenchymal stem cells, and thus a new generation beta cell differentiation method is developed. 3D bioprinting; The important advantages of the method are that it allows cells to be placed in the printed tissue while the tissue scaffold is still at the printing stage, that the produced tissue scaffold can be adjusted to the desired composition and can be designed according to the desired micropatterns, and it provides much more biocompatible niches than the matrices created by other methods. However, the material to be used in 3D bioprinting and the composition created are important. Since decellularized (non-synthetic, natural) material is used in the tissue scaffolds produced with this technique, it is very important that it offers the opportunity for bioprinting with the natural extracellular matrix of the organ. Because the ratios of extracellular matrix proteins such as collagen, fibronectin, laminin etc. in the tissue are preserved exactly, and thanks to the cells contained in the bioink, every region of the tissue allows easy cellularization. Extracellular matrix components are generally conserved structures across species and produce either no immune response or can be easily tolerated, even in xenogenic transplants. Thanks to this technique, both the natural structure is preserved and decellularized extracellular matrices obtained from different species can be used for treatment. Thus, the problem encountered in allogeneic and even xenogenic organ/tissue transplantations is overcome. The invention is also a cellular therapy product for T1D7. Endocrine pancreas engineering involves the development of new generation beta cell differentiation techniques and the production method of an endocrine pancreas-like functional tissue piece (biochip) that can bring blood sugar to a healthy level by secreting insulin in response to glucose. This "product", created by the development of many techniques, constitutes a new concept in itself. The problems related to the need for insulin in studies targeting only autoimmunity are overcome in this invention as follows: within the scope of the invention, the entire approach to the treatment of T1D7, both immunotherapy and beta-cell replacement, is presented. Preventing autoimmune attacks through in vivo programming of T regulatory cells (Treg) in individuals, 2) Creating an environment where autoimmunity can be prevented before biochip transplantation, so that the tissue-engineered biochip (insulin-secreting tissue), which is the basis of the invention, can continue to function in the in vivo environment, 3) By developing new generation beta cell differentiation techniques in endocrine pancreas engineering, the invention enables the production and transplantation of endocrine pancreas-like functional tissue parts that can bring blood sugar to a healthy level by secreting insulin in response to glucose. While differentiation is carried out, dendritic cell- and/or antigen-conjugated biodegradable nanoparticle-mediated Tregderin reregulation is provided to achieve immune tolerance against beta cells of autoreactive immune cells in T1D. Therefore, the invention can target the treatment of TlD disease as a whole. Thus, a new strategy that enables the treatment of TlD disease is provided by the production of personalized tissues and the tolerance of autoimmune T cells. Industrial Application of the Invention: The product produced within the scope of the invention has the potential to significantly prevent the expenses for commercially sold insulin, as it is a living tissue that can be produced specifically for the patient and can secrete insulin. In addition, since other complications seen in chronic periods in diabetic patients can be prevented, expenses for the treatment of these diseases will be reduced and perhaps eliminated. As a result, this product we have produced has a great potential to prevent a significant economic loss spent on diabetes. The production of the product is a suitable method for biofabrication. Personalized tissues can be produced for each patient by changing only the patient's cells. If the clinical use objectives of the method presented in the present invention as a treatment are achieved; This method will not only eliminate the economic burden required for the treatment of diabetes, but will also prevent many secondary complications that occur in the chronic period before they develop. Thus, it will bring high added value to our country. The target audience of the invention is TlD patients, but although the invention is a product created for the treatment of TlD disease, it will also serve as a model for the treatment of many other autoimmune diseases. The invention combines both beta cell replacement and immunotherapy, which are important approaches developed to date, with the current techniques of tissue engineering, and while doing this, rearranges the existing tissue engineering techniques to be suitable for clinical use, resulting in a brand new and very complex approach consisting of many steps. is the product. It is a multidisciplinary study that can be given as an example of studies in both the stem cell field, immunotherapy field, tissue engineering field and TlD field. TR