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 NEW GENERATION PANCREATIC TISSUE THAT CAN AVOID IMMUNE ATTACKS AND SECRETING INSULIN AND ITS PRODUCTION METHOD Technical Field The invention is related to the production of glucose-sensitive insulin-secreting tissues by developing new techniques in T1D (Type 1 Diabetes) and a therapeutic method that provides reprogramming of the immune system that causes autoimmunity and endocrine pancreas engineering. Previous Technical Field Type 1 diabetes is an autoimmune disease and the death of the beta cells in the body or their destruction to the extent that they cannot produce insulin is caused by autoimmunity involved in the pathogenesis of the disease [1]. Type 1 diabetes mellitus is a disease characterized by insufficient insulin production or complete inability to produce insulin as a result of autoimmune attacks that occur with the attack of autoreactive T cells on insulin-producing cells [3- (T1D) [2] is one of the diseases for which intensive efforts are being made today. The discovery of insulin has been a significant development for diabetic patients. Insulin, which cannot be produced in sufficient quantities by the pancreas, has begun to be supplied exogenously. For this reason, T1D is also called insulin-dependent diabetes mellitus (IDDM). Although T1D patients can maintain their lives thanks to the insulin they receive externally by injection, low quality of life remains a problem even after 80 years. Therefore, the solution to the disease was seen as enabling individuals with T1D7 to produce insulin again. Previously, pancreas transplantation, insulin-producing beta cell transplantation, and pancreatic islet transplantation were tried in patients. Thanks to developing technology and increasing knowledge about stem cells, beta cell transplantation, especially from mesenchymal stem cells (MSCs), has been developed. Differentiation and transplantation into individuals with TLD have been attempted. Gene cloning technologies have been utilized to increase the differentiation efficiency of stem cells and to better demonstrate beta cell properties, and various genes have been transferred to these cells [3]. The subsequent emergence of 3D cell culture techniques, which offer the opportunity to better mimic the body, has also influenced TLD studies, and researchers have attempted to obtain tissues capable of producing insulin. Every tissue in the body has a specific biocomposition that allows cells to perform their functions perfectly. Although the production of organ matrices using 3D printing has become quite a phenomenon in recent years, the biocomposition of the tissue scaffold to be produced is quite different from the natural composition of body tissues. Therefore, capturing the content in question in tissues printed with 3D bioprinters is crucial for ensuring that the cells cultured in the material used perform their correct biological functions and The subsequent translation of the produced tissues into clinical practice is an important issue that remains to be resolved. One of the recent innovations has been the cultivation of islet-like structures on decellularized liver extracellular matrix and the attempt to create 3D tissues capable of secreting insulin [4]. Furthermore, the reshaping of the extracellular matrix obtained after decellularization by using it as a raw material in SB-bioprinters is a newly emerging technique. Adipose, cartilage, and heart tissue templates have been tested in vitro [5] and only one study involving heart tissue production involving transplantation into animals [6]. All these alternative TLD treatment approaches include beta-cell replacement, which can reverse the outcome of the disease by replacing the destroyed beta cells in the diabetic pancreas. However, aggressive autoimmunity persists even after beta-cell replacement. If this is not prevented, it is noteworthy that specific immune attacks will continue against newly transferred beta cells. Therefore, beta cell replacement alone is insufficient without eliminating the existing autoimmunity against beta cells. In many studies, Treg induction has been performed in vivo or in vitro to create Treg-based therapy approaches for autoimmune diseases. 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 are weaker than antigen-specific Tregs when transplanted to target autoimmunity [7]; [8]; A promising approach regarding polyclonal Tregs is the induction of the Treg population by low-dose IL-2 administration. The aim of this study is to increase the immune response and to establish tolerance in vivo [15]; [16]; [17]. However, the use of polyclonal Tregs, which are propagated in vitro or in vivo, carries the potential to induce full immune suppression, in other words, the risk of suppressing the beneficial immune system. On the other hand, since autoantigen-specific Tregs do not provide the advantage of antigen specificity and thus suppress the entire immune system, the treatment of autoimmune diseases has taken its place in the literature. Many studies, including T1D7, where the vehicle for disease regression has been shown in mice with autoimmunity, have shown that islet antigen-specific Tregs are more effective than polyclonal Tregs in preventing the onset of the disease. Moreover, only the transfer of autoantigen-specific Tregs has initiated the onset of the disease. It can stop existing and ongoing diabetes. Recently, approaches for the production or induction of Tregs have emerged both in vitro and in vivo. One of the approaches for in vitro autoantigen-specific Treg production is viral TCR gene transfer to T cells. Stimulation of naive CD4+ T cells transduced with TCR and/or FoxP3 gene in the presence of TGF-β 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 concentrations of IL-2 for the expansion of Tregs in vitro. Limitations in these applications have directed researchers to in vivo Treg induction. Therefore, although TGF-β-mediated induction of Tregs from naive T cells in the periphery is a frequently used method, this It has been determined that the suppression capabilities of injected Tregs are not permanent [26]; [27]. In preclinical studies on animals with T1D, the induction of autoantigen-specific Tregs was achieved with dendritic cells loaded with antigen in vivo [28] or in vitro [29]. Based on many studies demonstrating that antigen-specific tolerance of dendritic cells is mediated by Tregals, clinical trials have been conducted for the tolerance-based treatment of autoimmune diseases, including T1D [30], and its safety for T1D has been tested in a phase I clinical trial [31]. A new therapeutic trend has recently been developed in which the treatment of autoimmunity and transplantation immunity can be targeted by conjugating autoantigens to splenocytes or nanoparticles. [28]; [32]; [33]. In antigen-splenocyte or antigen-nanoparticle therapy, autoantigens are covalently bound to the surface of splenocytes or nanoparticles using ethylene carbodiimide (ECDI). This method has been extensively developed in animal models and a Sunday 1 clinical study on multiple sclerosis has been conducted [34]; [35]; [36]. Poly(lactic-co-glycolide) (PLG) is one of the most commonly used nanoparticles in this technique due to its biocompatible and biodegradable properties. It has been reported that when PLG conjugated with antigens via ECDl or splenocytes are transferred into living organisms, ECDl induces apoptosis and therefore the mechanism of antigen-splenocyte or antigen-PLG therapy is through the uptake of TGF-beta by phagocytes and apoptotic cells. This approach is thought to be dependent on the production of phospholipids [37]; [38]. This approach has been shown to be significantly more effective in the prevention and treatment of autoimmunity in various animal models compared to therapies using only soluble antigens or broad-spectrum immunosuppressive drugs without the use of splenocytes or PLG nanoparticles [32]. Induction of 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]. Even if the damaged tissue is restored by targeting Tregs with the effective mechanisms mentioned above, the autoreactive effector T cells present in treated individuals may exhibit resistance, and these remaining cells may cause disease relapse [39]. Therefore, for long-term treatment of autoimmune diseases, the existing autoreactive immune system in the patient must first be eliminated and Immune tolerance must then be achieved. A study and a patent for this study presenting a method for tolerizing the immune system of a person suffering from autoimmune disease demonstrate that the defective immune system is first destroyed in animal models of autoimmunity and then tolerance to autoantigens is restored by stimulating antigen-specific regulatory T cells. In this regard, inducing apoptosis in immune cells with antibodies or low-dose gamma irradiation to destroy the defective immune system, and then administering phagocytes with disease-related autoantigens for autoantigen-specific Treg induction will not be sufficient because the deficiency in the number of insulin-secreting cells in response to glucose as a result of beta cell destruction is another problem requiring a solution. Technical Problems in Current Practices and Technical Solutions Presented within the Scope of the Invention. Type 1 diabetes is a disease that is resistant to insulin-producing ß-cells in the pancreas. It is a disease that causes beta cell failure and subsequent disturbances in blood sugar control as a result of the development of an autoimmune response. Immunotherapy, in terms of treating T1D, addresses the root cause of T1D by resetting the balance between autoimmunity and regulatory mechanisms. T regulatory cells play an important role in this immune response [41]. An alternative T1D treatment includes beta cell replacement, which can reverse the outcome of the disease by replacing the destroyed beta cells in the diabetic pancreas. However, it is noteworthy that if aggressive autoimmunity persists after beta cell replacement, specific immunological attacks against the newly transferred beta cells will continue. Therefore, beta cell replacement alone, without eliminating the patient's existing autoimmunity against beta cells, is insufficient. On the other hand, immunotherapy for individuals with T1D This alone will not be sufficient because the deficiency in the number of insulin-secreting cells in response to glucose, resulting from beta cell destruction, will remain a problem requiring a solution [42]. Attempts to eliminate the autoimmunity problem of T1D with immunotherapy (dendritic cell or antigen-conjugated biodegradable nanoparticle-mediated Treg induction) are already existing approaches [43]. However, these studies only target immunotherapy. Although many approaches have been tried to replace damaged β-cells in T1D before, three of these strategies have come to the fore: (i) cell transplantation, (ii) islet transplantation, and (iii) pancreas transplantation [44]. However, pancreas and islet transplantation are both limited in terms of insulin supply due to difficulties in finding donor organs and serious problems because they require lifelong suppression of the immune system. The latter condition is treated with immunosuppressive drugs. However, the use of immunosuppressive drugs significantly hinders the functioning of the immune system, and these individuals face significant disadvantages in the fight against microbial diseases. Both problems have been solved by differentiating beta cells from autologous MSCs taken from patients with TLD and obtaining insulin-secreting tissues. Insulin-producing cells have been obtained by obtaining insulin-secreting tissues using the patient's stem cells (autologous MSCs) used within the scope of the invention. Since the cells that make up the tissue transplanted to the patient are the patient's own cells, no immune response develops against them, eliminating the need for immune system suppression. Type 1 diabetes mellitus is currently a disease with serious effects worldwide. The discovery of insulin is an important development for diabetic patients. There has been progress. Insulin, which cannot be produced sufficiently by the pancreas, has begun to be supplied exogenously. For this reason, insulin-dependent diabetes mellitus (IDDM) is also called insulin-dependent diabetes mellitus (IDDM). While patients with DIDM can maintain their lives thanks to insulin administered externally via injection, a low quality of life remains a significant problem even after 80 years. This problem has been solved by transplanting tissue fragments that can secrete insulin according to blood glucose levels. After the transplant, patients no longer require exogenous insulin. Cell transplantation, islet transplantation, and pancreas transplantation all raise questions about the migration of cells into the desired region, how many will migrate, which cell type to choose, and where the application site will be. However, the method we use The tissue scaffold we created not only creates a microenvironment for cells but also serves as a vector for them. This allows cells to be transferred to the desired site in the patient. The production of organ matrices using 3D printing has become quite popular in recent years. However, the biocomposition of the tissue scaffold to be produced is quite different from the natural composition of body tissues. Every tissue in the body has its own unique biocomposition. This composition has a special design that allows the cells in the tissue to perform their functions in an inefficient manner. However, capturing this content in tissues printed with 38 printers poses a significant challenge. However, the decellularized pancreatic matrix used in this invention as an organ printing material in 3D printing preserves the tissue's natural biological content. This approach, which can play a vital role in ensuring that cells cultured in the material perform their correct biological functions, also overcomes several extremely important challenges. Generating glucose-sensitive insulin-secreting beta cells from stem cells is a very difficult process due to low efficiency. Obtaining human beta cells suitable for transplantation is also very difficult. While viral methods offer the highest efficiency in gene delivery to cells to confer desired phenotypic and functional characteristics or to stimulate them in a specific direction, viral methods are not suitable for clinical use due to safety issues such as the risk of mutagenesis and immunogenicity. On the other hand, non-viral gene delivery vectors offer a safer profile in tissue engineering. However, expression of the transferred gene in the target cell has not reached the same level with non-viral methods. This demonstrates the importance of vector design. An ideal gene delivery vector for tissue engineering should be biocompatible, biodegradable, minimally cytotoxic, and capable of effectively delivering DNA into the cell. It should also be able to maintain expression of the target protein for the required period. Its use together with its scaffold has led to the birth of a new field: Gene-Activated Matrix (GAM). GAM is the direct and continuous delivery of nucleic acids from a tissue scaffold to cells to provide effective and durable non-viral gene transfer to the cell. Generally, the nucleic acids to be transferred to cells (a plasmid DNA, usually called pDNA, carrying the target genes) are encapsulated in cationic polymers or cationic lipids and mixed with the solution in which the scaffolds will be formed. SB scaffolds are then formed from materials that have acquired gene transfer properties, and these structures are called GAM. When cells are seeded 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]. Considering all the desired properties related to biocompatibility and gene transfer, the cationic polymer chitosan is thought to be one of the most suitable non-viral methods for this purpose. In particular, the development of a low molecular weight, water-soluble oligochitosan molecule is a valuable resource. In this invention, the problems related to generating beta cells that can secrete insulin sensitively to glucose levels from stem cells were solved by obtaining a gene-activating matrix from the natural pancreatic extracellular matrix transferred with chitosan-pDNA nanoparticles and by creating a tissue scaffold by printing this matrix in 3D bioprinters. Thus, the PdX-1 and MafA [47] genes, which play important roles during pancreatic development and induce ß-cell differentiation, were transferred to MSCs cultured on the scaffolds. By stimulating ß-cell differentiation from MSCs using gene-activating matrices generated in this invention, a new generation beta cell differentiation method for TLD has been developed. Technical problems in beta cell differentiation in existing methods have been overcome by providing the GAM-mediated 3D microenvironment produced within the scope of this invention. One of the most difficult problems to overcome in TLD to date has been autoimmunity against insulin-producing cells. To address this problem, cells have been encapsulated using encapsulation techniques, preventing direct contact with autoreactive T lymphocytes. However, these methods have not enabled long-term treatment. Therefore, autoreactivity remains a hot topic even after encapsulation. However, within the scope of this invention, we utilized dendritic cell- and antigen-conjugated biodegradable nanoparticle-mediated Treg programming, resulting in specific tolerance to autoantigens and subsequent implantation of a biochip containing insulin-producing cells. Thus, this problem was overcome by immunotherapy mediated by antigen-specific tolerance in autoreactive T lymphocytes. The low frequency of Tregs in peripheral blood has been identified in recent years as a major challenge in Treg-based immunotherapy, and approaches for their induction have emerged. One approach for generating autoantigen-specific Tregs in vitro 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 the transgenic TCR manipulation. Difficulties in determining the antigen specificity of Tregs in the production of in vitro autoantigen-specific Tregs and in obtaining sufficient numbers of antigen-specific Tregs are other issues that limit their clinical applications. These limitations have led researchers to in vivo Treg induction. In this regard, in vivo autoantigen-specific Tregs have been induced from naive T cells in the periphery via TGF-beta [27]; [14] or by antigen-loaded dendritic cells [28]. However, the fact that these studies were conducted before autoimmune diseases had developed prevents the therapeutic efficacy of these applications from being determined. Antigen-PLG or splenocyte therapy approaches for the prevention and treatment of autoimmunity have been shown to be significantly effective in various animal models compared to therapies using only soluble antigen without PLG nanoparticles or splenocytes, or broad-spectrum immunosuppressive drugs [32]. Induction of autoantigen-specific Tregs in this manner is a highly effective approach with exciting preclinical results in multiple sclerosis models and Type 1 diabetes models [37]. On the other hand, in disease states, immune cells that respond to autoantigens are uncontrollably activated by increased pro-inflammatory cytokines, causing Tregs to lose their suppressive capacity. Tregs can even transform into effector cells such as Th17 cells in this pro-inflammatory environment [24]. Even if damaged tissue is restored by targeting Tregs with the effective mechanisms mentioned above (such as antigen-PLG and dendritic cells), the autoreactive effector/memory T cells present in treated individuals may exhibit resistance, and these remaining cells may cause disease relapse [39]. To solve this problem and ensure long-term treatment of autoimmune diseases, the existing autoreactive immune system in the patient must first be eliminated, and then immune tolerance must be achieved. According to data from the International Diabetes Federation, diabetes-related mortality is expected to rise to 1,000 in 2015. T1D is a disease characterized by the attack of autoreactive T lymphocytes on insulin-producing pancreatic ß cells, resulting in insufficient insulin production. Therefore, patients with T1D must take commercially available insulin daily to meet their insulin needs. Furthermore, chronic T1D can lead to serious secondary complications such as eye damage, neuropathy, nephropathy, and cardiovascular disease. From this perspective, diabetic patients also face significant economic burdens related to other diseases caused by diabetes. The document describes a method for preparing autologous dendritic cells derived from mesenchymal stem cells. During the generation of dendritic cells capable of providing immunosuppression, mesenchymal stem cells were used as a stimulator, or supportive stimulus, to induce the dendritic cells to acquire their immunosuppressive function. To stimulate the dendritic cells to stimulate the mesenchymal stem cells, these two cell types were co-cultured. Among the known applications in the art, Chinese patent document CN105670990 describes a tissue engineering application and preparation method for improved directional differentiation from mesenchymal stem cells. The invention was made in the field of biomedical engineering and encompasses the preparation and application of a tissue engineering material to stimulate the differentiation of mesenchymal stem cells. The polymer used is poly(lactic-co-glycolic acid), called PLGA, which is classified as unnatural polymers, meaning polymers that are not produced or found in the body [49]. Although biocompatible, this polymer, which is not produced in the body, is a completely foreign environment for cells. As is known, organs consist of two components: cells and the intercellular material called the extracellular matrix [50]. In the decellularization technique used within the patent, cells are detached from the extracellular matrix, and only the natural extracellular matrix of the organ or tissue is obtained. Thus, the polymers used are a native medium of the organ already present in the body, obtained after organ decellularization. They go far beyond being composed of a single polymer like PLGA and include collagen type 1, collagen type 4, fibronectin, glycosaminoglycans, etc. It consists of a natural biomaterial with a complex content containing many natural polymers [51]. Replacing dead beta cells is not a permanent and long-term solution for the treatment of T1D. This is because autoimmunity against beta cells still persists in the body. Consequently, a permanent solution for T1D requires not only correcting the beta cell deficiency but also eliminating the existing autoimmunity. However, patent no. CN105670990 only targets beta cell replacement. Patent no. CN105670990 has not been shown to reduce high blood glucose levels to normal after transplantation of the produced beta cells into a diabetic individual, and no in vivo (in a living organism) studies are included. In short, the function of the product in question has not been demonstrated in vivo. The document describes the development of autologous and allogeneic adipose tissue-derived mesenchymal stem cell compositions for the treatment of diabetic wounds. Diabetic wounds are secondary vascular complications that develop in individuals with diabetes due to excessively elevated blood glucose levels as a result of the disease. Wound healing focuses on improving the vascular system (vessels) and inflammation. Autologous mesenchymal stem cells have been used for wound healing and inflammation prevention. A known property of mesenchymal stem cells is their ability to inhibit (stop, prevent) immune responses in the recipient area [52]. Mesenchymal stem cells have high potential for use in current clinical therapies, particularly due to their easy harvesting from many tissues, anti-apoptotic and anti-inflammatory properties, lack of immune response in allogeneic transplants, and ability to differentiate into many somatic cells, including ß-cells. Within the scope of the invention, a biodegradable tissue scaffold is transplanted into the human body. Among the known applications, Chinese patent document CN1O4353l 15 discloses a kit for pancreatic decellularized tissue scaffolds, their preparation method, and a method for reseeding the scaffold. The scaffold used in this invention is an extracellular matrix obtained from decellularized pancreas. The permanent solution for TLD is not only to correct the beta cell deficiency but also to target the existing autoimmunity by replacing only beta cells, as is the case with a few patents. However, the literature indicates that this cell line is not a pure beta cell line [53]. More importantly, however, this cell line is an insulinoma cell line. In other words, it is a cell line derived from a tumor originating from the B cells of the pancreatic islets of Langerhans. The information in question is also available at 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, it has not been shown that the high blood glucose level is reduced to normal values after the transplantation of the produced beta cells to a diabetic individual, and it is not included in any in vivo (in a living organism) study. In short, the function of the product in question has not been demonstrated in vivo. In the document, cell stimulation biochip and detection of differentiation of stem cells are mentioned. Among the applications known in the art, the United States patent document numbered USZO13017l75 describes the use of activated mesenchymal cells for wound healing and regeneration of damaged tissue. stem cells are mentioned. The invention is a tissue repair method that activates the immunosuppressive properties of mesenchymal stem cells via inflammatory cytokines, thus initiating wound healing and angiogenesis (vascularization). The aim is to demonstrate how effective the use of mesenchymal stem cells with activated immunosuppressive properties can be in preventing inflammation caused by graft-versus-host disease (GVHD), which develops as a result of rejection of the transplanted tissue by the recipient in tissue transplants. In this regard, the initiation of the activated mesenchymal stem cell composition created for the prevention of GVHD in vivo involves the generation of a composition of mammalian mesenchymal stem cells activated with inflammatory cytokines to inhibit inflammation and promote tissue repair, and the transplantation of the cells to this site. This The basic methodology of the invention is the use of immunomodulatory properties of mesenchymal stem cells in the inflammatory environment. It is claimed that their use to prevent islet transplantation may also be functional. In the invention numbered USZO130l7l75, the cell composition applied in vivo for tissue regeneration and organ repair consists of mammalian mesenchymal stem cells activated with inflammatory cytokines. The application document includes an immunotherapy and kit developed for the treatment or tolerization of autoimmune diseases expressed by apoptosis-antigen therapy. The therapeutic steps of the invention are as follows: i) Identification 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 applying low-dose radiation (radiotherapy); (iii) by administering macrophages to the patient following radiotherapy or antibody administration; and (iv) finally, by administering autoantigens that cause autoimmunity to the patient. As a result of all these steps, existing autoimmune cells in the radiotherapy area were eliminated by inducing apoptosis, and the administered macrophages were able to support stronger immune tolerance when they encountered apoptotic cells. After this environment was established, an immunotherapy approach that stimulates Tregs by administering autoantigens associated with autoimmune disease to the patient, a common method of antigen-specific therapy, has recently been presented. The common aspects found when both patents were evaluated were that, in the immunotherapy part applied for TLS, Tregs are transferred to patient individuals in vivo via antigen-specific macrophages. In the present invention, autoantigens are not administered to patient individuals, but rather are loaded (stimulated) onto dendritic cells in vitro to produce antigen-specific dendritic cells and/or conjugated to biodegradable nanoparticles (PLGA-Antigen). As a result, the produced antigen-specific dendritic cells and/or PLGA-Antigen nanoparticles are administered to the patient, enabling the Tregs in the immune system to be programmed specifically for the antigens we selected. The purpose of the invention is only immunotherapy, immunotherapy applied to individuals with T1D will not be sufficient alone because the deficiency in the number of insulin-secreting cells in response to glucose as a result of beta cell damage will remain a problem requiring a solution. Brief Description of the Invention This invention is a T1D treatment method that aims to eliminate the existing autoimmunity in T1D patients through reprogramming and to provide insulin-producing autologous cell replacement. In line with this purpose, the invention, on the one hand, prevents the autoimmune immune system that attacks the insulin-producing ß-cells in T1D through in vivo programming of T regulatory cells (Treg). On the other hand, it provides the creation of an environment in which autoimmunity can be prevented so that the produced tissue-engineered biochip (insulin-secreting tissue) can maintain its function in vivo before tissue transplantation. provided. The invention also encompasses the production method of a functional endocrine pancreas-like tissue fragment capable of regulating blood sugar levels by secreting insulin in response to glucose, by developing next-generation beta cell differentiation techniques in endocrine pancreas engineering. In this respect, the invention encompasses both a product and a production method for this product. The aim is to prepare the components that constitute the insulin-secreting tissue (mesenchymal stem cells, beta cell gene delivery system, and decellularized pancreatic matrix (dpESM)), to obtain a tissue prototype by printing these components using 3D bioprinters, and to culture this tissue prototype in vitro. Thus, the stimulation of endocrine-pancreatic differentiation of mesenchymal stem cells is achieved through the influence of many factors [(i) dpESM, (ii) the gene transfer system related to ß-cell differentiation, and (iii) endocrine medium culture]. The invention also includes a 3D bioprinted endocrine pancreas-like functional tissue part product, the Gene Activating Matrix (GAM; dpESM + cationic polymer-pDNA nanoparticles), which consists of decellularized pancreas integrated with a cationic polymer-pDNA nanoparticle gene transfer system suitable for differentiating mesenchymal stem cells into beta cells. This endocrine differentiation-stimulating GAM is a sub-product created within the scope of the invention and is one of the subjects of the invention. In the invention, on the other hand, the produced biochip (insulin secreting tissue) is transplanted to an individual with T1D whose autoimmune attacks have been prevented. After all these processes, the ultimate goal of the study is to reduce the blood sugar to normal levels thanks to the biochip transplanted to T1D patients whose autoimmune attacks have been prevented. Detailed Description of the Invention One subject of this invention is that it is a therapeutic method for T1D disease. The present invention provides a treatment method that provides ß-cell regeneration in T1D by means of 3D culture environment while also reregulating autoimmunity. While ß-cell differentiation is carried out from MSCs on the tissue scaffold produced in order to provide a microenvironment very close to the natural one, autoreactive T cells against these cells; This invention involves the dendritic cell- and/or antigen-conjugated biodegradable nanoparticle-mediated Treg re-regulation. Therefore, the invention can target the treatment of T1D disease as a whole. Thus, a new strategy is provided that allows the treatment of T1D disease by generating personalized tissues and ensuring tolerance of autoimmune T cells. Another subject of the invention is that it includes a new generation beta cell differentiation method in pancreatic tissue engineering. The tissue scaffold designed for B-cell differentiation is printed using natural decellularized pancreatic extracellular matrix containing a gene transfer system and this is the first demonstration of the applicability of this method. Thanks to this invention, the natural polymer structure of the pancreas-derived matrix (dpESM) in the tissue produced using 3D bioprinters is transformed into a non-viral ß-cell gene transfer system. The gene-activating matrices obtained in this way provide a suitable 3D microenvironment for beta cell differentiation, thus enabling the development of a new generation beta cell differentiation method. The invention, as a tissue engineering application and preparation method, primarily includes the following innovative steps: a) Transformation of the product obtained by lyophilization of decellularized pancreas (dp-ESM) into a material that can be printed on 3D bioprinters. b) Preparation of bioink (Bioink: dpESM + chitosan-pDNA nanoparticles + MSC composition). o Integration of a nanoparticle gene (β-cell differentiation-related genes) delivery system into dpESM using a non-viral, i.e., cationic polymer instead of a virus. o Chitosan-pDNA o Adding MSC composition at a certain concentration and under certain environmental conditions to the dpESM mixture containing the chitosan-pDNA nanoparticle ß-cell gene delivery system c) Printing Insulin Secreting Tissue (Biochip) from Bioink with a 3D printer Another subject of this invention is the creation of a treatment product for T1D. It is the production of a transplanted endocrine pancreas-like functional tissue piece (biochip) to help regulate blood sugar to healthy levels in individuals with T1D whose autoimmune attacks are prevented. This "product", created by the development of many techniques, constitutes a new concept in itself. The outline and content of the method used within the scope of the invention, which aims to produce endocrine pancreas-like tissue grafts suitable for lowering glucose-sensitive blood sugar after preventing autoimmune attacks in patients with T1D (Type 1 Diabetes), and to transplant them in the form of subcutaneous biochips, are as follows: - Patient-specific selection of T1D autoantigen (ß-cell antigen) combinations and doses to be administered as intravenous injections to T1D subjects after the T1D subjects are identified*, - Obtaining immature/naive dendritic cells from bone marrow progenitors or blood mononuclear cells of T1D subjects in order to provide antigen-specific tolerance in T1D subjects, - Autologous to induce tolerance in order to program the existing Tregs in the patient antigen-specifically. Immature/naive dendritic cells are obtained from bone marrow progenitors of T1D71i individuals by culturing them with GM-CSF cytokine. Briefly, bone marrow (KI) cells are obtained from femur and tibia sources and erythrocytes are subjected to hypotonic lysis. KI cells are cultured in 10 ml of medium supplemented with cytokine. Half of the cell medium is renewed daily with fresh medium containing cytokine. To determine the purity of dendritic cells, cells are labeled with CD110, a dendritic cell marker, on the 7th day of culture and analyzed by flow cytometry. The obtained dendritic cells are co-cultured with previously prepared lysate (apoptotic bodies of ß-cells) and GAD65 peptide in the next specified step. When dendritic cells phagocytose the apoptotic bodies and peptides, they are loaded with the targeted antigens. (primed/pulsed) term. - In order to ensure antigen-specific tolerance in TID subjects, the determined TID autoantigens are loaded into dendritic cells obtained from subjects in vitro and/or conjugated to synthetic biodegradable nanoparticles***, lysate (apoptotic bodies of IS-cells) is prepared, and ß-cells are apoptotic. First, the prepared ß-cell line or the physically homogenized 3B Biochip is seeded in 24-well petri dishes with 500 µl medium in each well at 3<105 cells/well. After 45 minutes of irradiation with ultraviolet B (UVB, 10 mJ/m2), it is cultured overnight in 5% CO2, 37°C. Characterization of apoptosis is performed using annexin Confirmed by V-fluorescein isothiocyanate (FITC) and propidium iodide staining in Ilow cytometry. Co-culture and characterization of dendritic cells with autoantigens (ß-cell antigens) The obtained dendritic cells were co-cultured with apoptotic bodies and GAD65 peptide as ß-cell specific antigens. For co-culture, 106 cells/well dendritic cells: 3x105 cells/well apoptotic beta cells (3:1 ratio) were cultured in 24-well plates. The prepared culture was continued for 2 hours by adding synthetic GAD65 peptide in SuM. Dendritic cells loaded (primed) with diabetogenic antigens (beta cell apoptotic bodies and GAD65 peptide) were characterized by flow cytometry analysis. Before co-culture, CFSE and after co-culture, CD11 were After analysis of labeled dendritic cells, dendritic cells loaded with apoptotic bodies were determined to be CD110- and CFSE-positive, while unloaded dendritic cells were determined to be CD11c+ and CFSE-negative. For the characterization of dendritic cells that will exhibit immune tolerance, after activation by the addition of liposaccharide to their cultures, CD40 and CD86 co-stimulator expression and the release of pro-inflammatory cytokines were analyzed. Dendritic cells that exhibited tolerance based on their resistance to the increase in co-stimulatory expression and the release of pro-inflammatory cytokines were characterized by flow cytometry and the medium was analyzed 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, were synthesized using ethylene carbodiimide (ECDI) As mentioned in the previous steps, ß-cell specific antigens were isolated ß-cell apoptotic bodies and conjugated with commercially available GAD65 peptide to form a biodegradable antigen-conjugated nanoparticle (antigen-ECDI-PLG nanoparticle). Briefly, ECDI [(1-Ethyl-3-(3' dimethylaminopropyl) carbodiimide HCl] was used to conjugate ß-cell specific antigens to biodegradable PLG nanoparticles in vitro. PLG particles (3 mg) were washed 3 times with PBS to remove sugars from lyotylization and incubated for 1 hour with 30 mg/ml ECDI and 1200 µg/ml lysate (ß cell apoptotic bodies) and 5 µM GAD65 peptide for each dose. The conjugated particles are washed twice with PBS to remove excess ECDF and passed through a 40 µm cell strainer. The efficiency of lysate and peptide conjugation is characterized by measuring the amount of protein remaining in the supernatant by protein assay. Before being converted into lysate, PLG nanoparticles are conjugated with CFSE-labeled GAD65 and CFSE-labeled ß-cell apoptotic bodies by flow cytometry. - Inducing apoptosis of existing autoimmune system cells in T1D patients with low-dose (supplemental level) radiation to induce antigen-specific tolerance. - Subsequently, dendritic cell and/or antigen-conjugated biodegradable nanoparticles are injected intravenously (IV) into T1D patients. ß-cell specific antigens (GAD65 peptide and lysate) are conjugated with ß-cell specific antigens (GAD65 peptide and lysate). For the application of loaded dendritic cells, approximately 2-3 million macrophage/dendritic cells/injection can be injected for each mouse and approximately 8-12 million macrophage/dendritic cells/injection for humans, and the dose and number of injections can be increased according to the patient's immune response. In antigen-conjugated-nanoparticle-mediated tolerance induction, each patient can be transplanted with antigen-loaded nanoparticles (approximately 10 µg of peptide-containing nanoparticle/injection for humans, a total of at least 30 µg, 1-2 µg/injection for mice), and the dose and number of injections can be increased according to the patient's immune response. - While the above process steps are being carried out, the pancreas is decellularized to obtain cell-free pancreatic extracellular matrix (dpESM). The process of obtaining decellularized pancreatic extracellular matrix (dpESM) by perfusion decellularization from the pancreas is carried out under sterile conditions in an airtlow cabinet (Heraeus Instruments, Hanau, Germany). In order to lyse the isolated pancreatic cells (dpESM, which does not develop an immune response, can be isolated allogenically or xenogeneically from the pancreas of rodents such as rabbits, rats, mice and other mammals, and human cadavers) and to remove the organ from the extracellular matrix, a retrograde (reverse direction) perfusion method is used via the hepatic portal vein connected to the perfusion system. For this purpose, first, 1% TritonX-100 is perfused through the portal vein at 10 ml/min for 60 minutes. Then, it is perfused for 120 minutes. Then, it is perfused again with 1% TritonX-1000 for 15 minutes. The matrix is perfused at 10 ml/min for 10 minutes. Sterilization of the matrix is achieved with 300 ml of 0.1% peracetic acid. The enzymes and chemicals are removed from the tissue by perfusing at 2 ml/min for 4 hours in phosphate buffer (PES) with a pI-Psi of 7.4. For dpESM characterization, firstly, the DNA content is determined by PCR method in dpESM. After the decellularization process, histochemical and immunofluorescence analyses are performed to determine whether the parenchymal and stromal integrity of the pancreas is preserved in dpESM, whether any cells remain, and also to determine the glycosaminoglycan (GAG) content and are compared with native pancreatic tissue. - Afterwards, the decellularized pancreatic matrix (dpESM) The product obtained by lyophilization and powdering is converted into a solution form material that can be printed in 3D bioprinters, and for the lyophilization and powdering of dpESM5, a lyophilizer (FTS Systems Bulk FreezeDryer Model 8-54) is used to completely remove the water retained by dpESM. Decellularized pancreatic tissue is kept wrapped in aluminum foil to prevent freezing at -200 C0, and all samples lyophilized in the lyophilizer for 20 hours (±2 hours) between 0 and -100CCJ are stored in airtight sealed packages. They are then sterilized by e-beam (electron-beam) irradiation at 22 kGy. The lyophilized sheets are immersed in 0.9% saline for 5 minutes. To powder dpESM, the lyophilized The resulting layers are crushed in a mill compressed with liquid nitrogen. dpESM powder is prepared by acid extraction of these particles at room temperature for 3 hours. It is then washed by centrifuging with sterile distilled water at 1000g at 4°C for 10 minutes. dESM powder is extracted with ethanol and ether, and the ether is evaporated under a chemical detonator. The protein content of dpESM is determined by LCMS-IT-TOF analysis. Dried dp-ESM is thoroughly crushed in liquid nitrogen with a mortar and pestle, and to solubilize it; it is mixed with 10% pepsin (W/W) prepared with 0.5 M acetic acid for 48 hours at room temperature. Ion balance is achieved using 10x PBS. The solution is dissolved in undissolved matrix particles. The final concentration of the solubilized dpESM is adjusted with 0.5 M acetic acid. Mycoplasma screening is then performed with commercially available kits (such as MycoAlert™ Myeoplasma Detection Kit). The pH of the dpESM solution is expected to be approximately 2.8-3 at this stage. This pH value is adjusted to 7.4 using 10 M NaOH. All these operations are carried out at 10 ° C to prevent the solution from gelling. It is kept in liquid form for the preparation of the bioink. Meanwhile, a cationic polymer (chitosan or other cationic polymers that are not toxic to the subjects) is used to transfer genes related to ß-cell differentiation in a non-viral way and Chitosan-pDNA nanoparticle ß-cell gene delivery system**** formation by ionic gelation method. Chitosan-pDNA nanoparticles to be used as gene delivery system are commercially available as oligochitosan (MW 7.3kDa; DD 97%) molecule (Novamatrix, FMC Biopolymer, Norway). Chitosan-pDNA nanoparticles are formed by the ionic gelation formula; electrostatic interaction between cationic (+) chitosan and anionic (-). In summary, chitosan is dissolved in 1%-2% acetic acid and added dropwise to a continuously stirred aqueous solution containing pDNA and sodium tripolyphosphate (TPP) to form cross-links between chitosan-pDNA particles. Chitosan-pDNA nanoparticles precipitated as a result of ionic gelation are It is kept at room temperature for 30 minutes for stabilization before use. Gene Activating Matrix (GAM; dpESM+chitosan pDNA nanoparticles) is obtained 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 in a 3D bioprinter. Obtaining autologous mesenchymal stem cells (MSCs) from bone marrow or other tissues and preparing MSCs for the production of endocrine pancreas-like tissue (biochip) in terms of number and phenotype for endocrine differentiation. For the isolation of bone marrow (BM)-derived mesenchymal stem cells (MSCs) from individuals with TLD, bone marrow aspirates are collected into heparinized tubes under appropriate conditions. In a sterile cabinet, bone marrow aspirates are washed twice with HBSS buffer (Hans Balanced Salt Solution) by centrifuging at 300g. They are then placed in 0.81% ammonium chloride and incubated at +4C for 15 minutes. After centrifugation and washing with PBS, the culture medium is diluted with DMEM-FIZ (Dulbecco's modified Eagle's medium-low glucose and F12 supplement) containing ng/mL bFGF and 1% glutamax. They are plated in flasks and cultured in an incubator at 37°C and 5% CO2. The medium is changed every 3 days, and when the cells are 70-80% confluent, adherent cells are removed by trypsinization and passaged at a 1:3 ratio. The cells obtained at the end of the 3rd passage are characterized. MSCs obtained from bone marrow tissue of individuals with TLD were isolated thanks to their adherence to culture plates, and the morphological characteristics of the adherent cells were examined using a contrast-phase microscope throughout the study. Flow cytometric analysis (MSCs surface markers for CD73, CD90, and CD105 positivity and CD45/CD34 negativity) and immunocytochemical labeling (Immunohistochemical Labeling (IHC) and Immunofluorescence Labeling (IF)) were performed to determine immunophenotypic characteristics. Gene expression was determined by RT-PCR. - Bioink is prepared by adding chitosan-pDNA nanoparticles and MCHs to the printable dpESM form - by providing appropriate conditions to prevent gelation -. After the previously mentioned technique, which is carried out at 10°C to prevent gelation of the dpESM solution, the following elements are added to the dpESM, which is kept in liquid form to obtain the bioink: Chitosan-pDNA nanoparticles; The chitosan-pDNA nanoparticles prepared as previously mentioned and the prepared dpESM solution are gently mixed. MCHs.' To prevent damage to cells and DNA-containing nanoparticles 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 purpose, before printing, the gelatin methacryloyl (GelMA) (10% (w/v) and 0.5% (w/v) 1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl-1-propanone photoinitiator (PI) solution is added to the cell count at a concentration of 5x105 cells/ml. Bone marrow-derived MSCs from the 3rd passage (P3) are removed from the culture vessel and added to the mixture of dpESM+chitosan pDNA nanoparticles at a concentration of 6x106 cells/ml and mixed gently. With the addition of cells, the dpESM+chitosan pDNA nanoparticles+MSCs composition is called bioink. - Printing the biochip from the bioink with a 3D printer, A high-resolution (~1.25-50 micron) multi-head 3D bioprinter, an extrusion-based bioprinter, with adjustable heating and cooling features is used for the printing of 3D tissue scaffolds designed for the differentiation of MSCs into pancreatic ß-cells. Hypoprinting is performed at 25°C, 25°C, 27.5°C, and 30°C using temperature control to optimize cell viability and material printability. The results are evaluated to determine the optimum conditions for each gel. Following bioprinting, the generated tissue scaffolds are cross-linked and gelated by applying UV light at 850 mW for 17 seconds from a distance of approximately 8.5 cm to the printed tissue product to ensure rapid cross-linking. (photo-polymerization by photocuring). In cases where the cross-linking process cannot be achieved at the desired degree, alginate is added in the same amount as GelMA added to the bioink (V/V) and after printing, it is left in 10ÜmM CaC12 solution for 3 minutes. Cell proliferation/viability analysis of the biochip obtained from the bioink with a 3D printer Determination of cell viability in the biochip is determined by two separate assays: 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] 3D cell culture viability analysis kits (as in the 3D Cell Culture HTS Cell Viability Complete Assay Kit, Biovision commercial product). - SB from bioink By culturing the bioprinted tissue in an endocrine differentiation medium, insulin-secreting beta-like cells are obtained from MSCs present in the tissue. The differentiation of MSCs into ß-cells within the biochip is induced physically thanks to the tissue scaffold obtained from the pancreatic extracellular matrix, induced at the genetic level by the MafA and Pdx-l transcription factors carried by the gene-activating matrix included in the biochip design, and induced by adding inducing factors to the medium. Thus, the ß-cell differentiation of MSCs is supported more strongly. For pancreatic ß-cell differentiation, MSCs are induced with a four-stage protocol. Differentiation is first initiated with 10% Human AB Serum (if the patient is human, fetal calf serum is used for other mammalian patient individuals), 10 mM nicotinamide, and 4 The cells are initiated by culturing in LG-DMEM medium supplemented with 25 ng/ml recombinant EGF and 0.5 mM ß-mercaptoethanol for 3 days. Then, the cells are incubated in LG-DMEM medium containing 25 ng/ml recombinant EGF in the second stage for 5 days. In the third induction stage, the cells are cultured in H-DMEM containing 2% Human AB Serum, 10 mM nicotinamide, 10 HM Exendin-4, 10 µg/ml INGAP-pp and 1)( ITS and recombinant bFGF and the differentiation is completed. The cells are incubated under controlled culture conditions of 37°C, 5% CO2. The media of the biochips are divided into two It is changed and refreshed once a day. In order to evaluate the differentiation of MSCs 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. In order to determine the secreted insulin level, INSULIN and C-PEPTIDE ELISA analysis (Human C-Peptide ELlSA Enzyme-Millipore) is performed and in order to see at the protein level, GATA-4, HNF3B, HNF4A, INSULIN I, INSULIN C-PEPTIDE, PDX-1, NKX2.2, MAFA, NGN3, MAFA and ISL-1 are immunofluorescently labeled. To characterize the biochip scaffold's similarity to the parenchymal and stromal structure of the native pancreas, Hematoxylin-Eosin (H-E), Alcian Blue, and Sirius Red stains are examined by histochemical methods. To determine the apoptotic damage status of the cells in the biochip, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) is performed using apoptosis detection kits (such as In Situ Cell Death Detection Kit, F. Hoffmann-La Roche). For the morphological analysis of the biochip, the insulin level in the biochip's medium is determined using a commercial kit (Millipore method). To determine whether the ß-cells in the biochip secrete insulin in vitro, depending on the glucose added to the medium, two The cells are exposed to different glucose concentrations and insulin is determined in the collected medium. Before starting the insulin secretion analysis, differentiated ß-cells are cultured for 48 hours in insulin-free medium and washed periodically with PBS until insulin is completely removed from the medium. Serum-free LG-DMEM (low glucose content; 5.5 mmol/L) containing 0.5% BSA is added to the wells and the cells are incubated for 1 hour at 37°C. The supernatant is collected for basal insulin secretion analysis and frozen at -70°C. Then, the cells are cultured in H-DMEM (high glucose content; 25 mmol/L) medium for another 1 hour. At the end of this period, the supernatants are collected and frozen to determine glucose-stimulated insulin secretion. Glucose-stimulated insulin levels and protein levels in ß-cells (experimental group), undifferentiated MSCs (negative control), and commercially available ß-cell line (positive control) secreted supernatants are determined in a microplate reader. - The study consists of the transplantation of pancreas-like functional tissue grafts as subcutaneous biochips to T1D subjects and the biochip's insulin secretion in response to glucose, thus controlling blood sugar to healthy levels. To determine whether there is bacterial contamination on the biochip after differentiation by conducting a pre-transplantation bioburden contamination analysis of the biochip, 5 ml of differentiation medium is taken into a 15 ml conical bottom tube and mixed with 7 ml SOC medium. Absorbance is determined at 600 nin. Following immunotherapy, biochips prepared in vitro are implanted subcutaneously in anesthetized individuals with T1D. Evaluation of the therapeutic effect of the transferred biochip: Individuals are monitored for 23 weeks after transplantation, and fasting blood glucose levels (FBG) are measured with periodic blood samples. FBG levels of 2126 mg/dl are considered diabetic. C-peptide and insulin ELISA analyses: Serum samples are collected on days 1 and 140, and human-specific C-peptide levels and human nonspecific insulin levels are measured using an ELISA assay kit (Wako Osaka, Japan and Shibayagi, Gunma, Japan). Oral glucose tolerance test: The function of the biochip to be transferred will be assessed with an OGTT (Oral Glucose Tolerance Test) on day 30. This test is performed by giving oral glucose solution to an individual with TLD who had been fasting for 8-12 hours (adults: 75g glucose solution, children: 1.75g/kg body weight) and measurements were made at different hours (0, 30, 60 and 120 minutes). AKS of 2 126 mg/dl and postprandial blood glucose of 200 mg/dl are considered diabetic values. Blood glucose measurements in diabetic individuals were used to confirm the presence of hyperglycemia (fasting blood glucose level of 2 126 mg/dl (11, postprandial blood glucose of 200 mg/dl) in individuals with TLD. For this purpose, a portable glucometer device (Medisense Precision QlD; Abbott Laboratories, Bedford, MA, USA) After 72 hours, blood samples are taken from individuals to determine blood glucose levels. The glycosylated hemoglobin (HbA1c) test is valuable in monitoring diabetic patients and provides an idea of the average blood glucose level over a three-month period. The ideal HbA1 level in diabetic patients is considered to be <6.5%. Consequently, a technique has been developed that prevents autoimmune attacks in patients with T1D as a result of tolerance-based immunotherapy. Furthermore, the transplantation of glucose-sensitive insulin-secreting tissue grafts normalizes blood glucose levels and maintains normal blood glucose levels for the long term. The most important feature of this technique in its therapeutic use is the ability of the transplanted tissue grafts to prevent autoimmune attacks and lower blood glucose in glucose-sensitive diabetic individuals.* Diagnostic testing or evaluation of individuals with T1D is performed using the oral glucose tolerance test (OGTT), glycosylated hemoglobin test, or fasting plasma glucose test. Furthermore, antibody testing is used to determine the type of diabetes antigen-reactive immune system in T1D patients. The information obtained from this is used to select personalized autoantigens. ** In the inventive method aimed at personalized treatment, instead of or in addition to GAD65, which is used as an autoantigen, antigen(s) detected at the highest levels by autoantibodies and identified in the development of diabetes can be used as biomarkers of ß-cell destruction in individuals with T1D. For example, nonspecific islet cell antigens (ICAs) insulin, insulinoma antigen-2 (lA-2), and transmembrane protein tyrosine phosphatase (lCA512). Islet-specific glucose-β-phosphatase catalytic subunit-related protein (IGRP) (Wenzlau and Hutton, 2013). The antigen to be selected will be determined by subjecting the individual with TLD to the antibody test used for the diagnosis of T1 diabetes, under the supervision of a physician or veterinarian with standard experience in this technique. Alternatively, loading the autoantigens onto dendritic cells obtained from the subjects and/or conjugating them to synthetic biodegradable nanoparticles) can be used. However, in cases where they are insufficient to prevent autoimmune attacks, they can also be used together to increase the immune tolerant effect. The choice of which method to use in the alternative use can be determined based on the availability of dendritic cells from patient subjects under the supervision of a physician or veterinarian with standard experience in this field. The autoreactive T cells that cause TLD are sensitive to β-cell antigens. To induce immunotolerance, tolerizing antigen-conjugated nanoparticles are obtained by in vitro conjugation of biocompatible and biodegradable nanoparticles (such as co-glycolide) with polymeric nanoparticles or liposomal nanoparticles [TlD autoantigens]. After the injection of antigen-conjugated nanoparticles into subjects with TlD, existing Tregs in the patient are reprogrammed towards tolerance. The pDNA contains Pdx-1 and MafA genes or transcription factors involved in cell development [3]. By combining chitosan and pDNA with ionic gelation, nano-sized chitosan-pDNA particles are obtained as gene transfer vectors. The term "individual" or "subject" when referred to as a subject or individual with TLD refers to any mammal, human or non-human. Dosage regimens for immunotherapy administered to individuals with TLD by intravenous injection 24 hours after radiation exposure can be adjusted to provide the optimal target response (e.g., tolerance induced in a subject and/or therapeutic effect). 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 decreased or increased according to the demands of the disease state. TLD autoantigens (ß-cell antigens); Ready-made synthetic antigens identified as autoantigens in T1D (GAD65 and/or 3B) are antigens obtained from lysates of insulin-secreting tissue/apoptotic bodies of ß-cells. In vivo antigen-specific programming of Tregs is carried out through the following method steps: i) Identification of autoimmune subjects; ii) Inducing apoptosis of immune system cells by application of low-dose radiation (radiotherapy); iii) Loading of determined T1D autoantigens onto dendritic cells obtained from subjects in vitro and/or conjugating them to synthetic biodegradable nanoparticles in order to induce antigen-specific tolerance in subjects with T1D; iv) Administration of autoantigen-loaded dendritic cells and/or antigen-conjugated biodegradable nanoparticles to the T1D subject following radiotherapy. An endocrine pancreas-like tissue piece (biochip) created with a 3D bioprinter obtained by the method in question contains pancreatic extracellular matrix obtained by lyophilization of decellularized pancreas, chitosan-pDNA nanoparticles gene (PdX-1 and MafA genes or [3 other transcription factors involved in cell development) delivery system and autologous (bone marrow-derived or obtained from other tissues) mesenchymal stem cells. The product created with a 3D bioprinter contains Gene Activating Matrix (GAM; dpESM+chitosan pDNA nanoparticles) consisting of decellularized pancreas integrated with chitosan-pDNA nanoparticle gene delivery system suitable for differentiating mesenchymal stem cells into beta cells. GAM, which stimulates this endocrine differentiation, is a subproduct 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 is created using chitosan or other cationic polymers that are non-toxic to the subjects, creating a chitosan-pDNA nanoparticle ß-cell gene delivery system. The method in question is for therapeutic use in the treatment of patients with T1D by using this endocrine pancreas-like tissue fragment (biochip). The product, implanted subcutaneously, lowers blood sugar by secreting glucose-sensitive insulin. The product obtained within the scope of the invention is suitable for clinical use because it uses a non-viral cysteine for gene transfer. Additionally, the product can be produced using 3D bioprinters according to the desired micropatterns after the matrix obtained by decellularization of the organ is pulverized and designed electronically using software. This allows for the integration of non-viral gene delivery systems into the matrix. The product is a matrix that allows for the addition of various polymers, growth factors, and media stabilizers to the solution obtained by solubilizing the decellularized pancreatic matrix, if needed. The inventive method allows for the redesign of the matrix in a computer environment, allowing for the final 3D shape of biochips, and for the matrix to be designed according to the target region to be transferred to the host. If cell viability is low in the sample tissue printed from the bioink used as a cellular matrix solution, a solution of the gene delivery system mixed with the matrix in solution can be used as a bioink, and printing can be achieved using these two elements in a 3D bioprinter. MKHWer can be added to the tissue scaffold after printing and during culture. A physician or veterinarian with standard experience with this technique can easily determine the effective dose of the components required for immunotherapy (dendritic cells, antigen-conjugated nanoparticles, or antigens). For example, to achieve the desired therapeutic effect through the immunotherapeutic components used, the physician or veterinarian initially administers the dose below the required level and gradually increases the dosage until the desired effect is achieved. One of the invention's subjects is the production of next-generation insulin-synthesizing tissue. A hybrid printing technique is developed by combining two distinct areas of tissue engineering, decellularization and αB bioprinting, to produce a bioactive matrix rewritten from the pancreas's own extracellular matrix using stem cells. Autologous mesenchymal stem cell differentiation yields insulin-producing beta cells, enabling the production of a functional, insulin-secreting, endocrine pancreas-like tissue fragment. Another subject of the invention is the therapeutic method for treating T1D, primarily by suppressing the autoimmune immune system that attacks the insulin-producing β-cells in T1D. After autoimmune attacks are prevented in individuals with TLD, insulin-producing tissue generated within the scope of the invention can be transplanted into individuals with TLD as subcutaneous biochips. Thus, the tissue can secrete insulin in response to glucose and bring blood sugar to healthy levels in diabetic individuals. Within the scope of the invention, patients with TLD are first identified, and the autoantigens and their doses to be administered to the identified TLD subjects are determined. To induce antigen-specific tolerance to TLD in subjects who develop TLD, antigen-loaded dendritic cells and/or antigen-conjugated biodegradable nanoparticles are generated. Both methods can be used as alternatives to induce antigen-specific tolerance in subjects with TLD. However, if preventing autoimmune attacks proves insufficient, they can also be used in conjunction with other therapies to enhance immunotolerance. The choice of alternative method can be determined by obtaining a sufficient number of dendritic cells from patient subjects under the supervision of a physician or veterinarian with standard experience in this field. Following apoptosis of existing autoimmune system cells in TlD patients with TlD using low-dose (supplethal) radiation, these dendritic cells and/or antigen-conjugated biodegradable nanoparticles are intravenously (IV) injected into the TlD patient subject, which then induces/reprograms T-regulatory cells (Tregs) to confer antigen-specific tolerance to TlD. Meanwhile, for the production of insulin-secreting tissue, which constitutes another part of the invention, the matrix that will form the scaffold of this tissue is obtained by pancreatic decellularization: acellular pancreatic extracellular matrix (dpESM), which is then lyophilized. The dpESM, which is completely dehydrated by lyophilization, is then ground into a powder and then converted into a solution at the appropriate concentration to maximize cell viability and enable bioprinting in 3D printers under optimal conditions. Meanwhile, as the second step in obtaining insulin-secreting tissue, a matrix is created that transfers ß-cell genes, enabling the scaffold to simultaneously transfer genes from mesenchymal stem cells (MSCs), which are used as cellular elements in the tissue, to insulin-secreting beta cells. Chitosan-pDNA nanoparticles, used as the ß-cell gene delivery system, are prepared by combining chitosan, a cationic polymer, with a pDNA vector carrying the ß-cell genes. Mesenchymal stem cells (MSCs), the final elements of the desired endocrine pancreas-like tissue, are also obtained at this stage and are prepared in terms of number and phenotype for endocrine differentiation. To prevent damage to the cells 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 the dpESM, which is kept in liquid form, and a pre-designed 3D tissue is printed using a 3D bioprinter. If 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 with the matrix in solution is used as bioink, and printing is carried out with these two elements in the 3D bioprinter. MSCs are added to the tissue scaffold after printing during culture. In both cases, the tissue scaffold formed after printing contains Gene Activating Matrix (GAM: dpESM + chitosan-pDNA nanoparticles). The 3D tissue obtained in both ways is in the endocrine differentiation medium. By culturing the mesenchymal stem cells in the tissue, they are differentiated in the endocrine direction and insulin-secreting beta-like cells are obtained. In this stem cell differentiation, a multi-faceted endocrine differentiation stimulation is provided thanks to GAM and endocrine culture medium. After the transplantation of endocrine pancreas-like functional tissue grafts (biochips) in the form of subcutaneous biochips to T1D patient subjects, the ability of the biochip to secrete insulin in response to glucose and to regulate blood sugar to a healthy level is tested. T1D subject or individual; the terms "subject or individual" mentioned refer to any of the human or non-human mammals that develop T1D. The element for which a product is created for the treatment of which and at the same time a therapeutic method is presented in the invention are T1D patient subjects. T1D 0t0antigens (ß-cell antigens); These antigens are designed to be loaded onto dendritic cells in vitro and/or conjugated to biodegradable nanoparticles to induce antigen-specific tolerance. These include: readily available, synthetically produced ß-cell antigens (such as GAD65) identified as autoantigens in T1D7; and antigens obtained from lysates of 3D tissue produced within the scope of this invention and from apoptotic bodies of ß-cells. Antigen-specific tolerant dendritic cells are generated by in vitro loading of dendritic cells obtained from the blood of subjects with T1D with T1D autoantigens. Antigen-loaded dendritic cells are generated to reprogram existing Tregs in patients with T1D toward tolerance. Dendritic cells are co-cultured with beta cell antigens that cause autoimmunity and are engineered specifically for the antigens that cause T1D autoimmunity, thus providing immunosuppression specific to these antigens. Within the scope of this invention, antigen-specific engineered dendritic cells are administered in vivo to stimulate T regulatory cells (Treg). Thus, to prevent autoimmunity in TLD, antigen-specific immunosuppression is achieved through in vivo programming of T regulatory cells (Treg) in an antigen-specific manner. Dendritic cells are loaded with autoantigens in vitro and made antigen-specific before being transferred to the patient. In other words, before dendritic cell transfer to the patient with TLD, dendritic cells loaded (stimulated) with autoantigens in vitro are generated. These antigen-specific induced tolerant dendritic cells are administered to the patients. Within the scope of the invention, biodegradable nanoparticles (PLGA-Antigen) to which the antigen is conjugated are used together with dendritic cells or as an alternative to dendritic cells. PLGA-Antigen nanoparticles are conjugated with selected antigens (beta-cell antigens) related to TLD. are being prepared. By administering PLGA-antigen particles to the patient after radiotherapy, Tregs in the immune system are programmed specifically for the antigens we have selected, thus offering an antigen-specific tolerance-based immunotherapy application. The antigens used in the invention (e.g., GAD65) are antigens derived from beta-cell apoptotic bodies (in combination if necessary depending on the patient). Antigen-conjugated biodegradable nanoparticles; biocompatible and biodegradable nanoparticles [polymeric nanoparticles such as PLG poly(lactic-co-glycolide), also known as PLGA poly(lactic-co-glycolic acid) or liposomal nanoparticles] are used to induce immunotolerance of autoreactive T cells that cause T1D against β-cell antigens in vitro with T1D autoantigens. Tolerant antigen-conjugated nanoparticles are obtained by conjugation. Antigen-conjugated nanoparticles are formed to reprogram existing Tregs in the patient towards tolerance after injection into T1D subjects. Low-dose radiotherapy; In subjects with T1D, existing autoimmune system cells are induced to apoptosis with low dose (supplemental level) radiation 1 s1n1m1. Radiotherapy is administered both to eliminate existing autoimmune cells by inducing apoptosis and to support immune tolerance more strongly when the antigen-loaded dendritic cells and nanoparticles administered after radiotherapy encounter apoptotic cells. In order to provide immune tolerance to ß-cell antigens of autoreactive T cells that are the cause of the disease, antigen-loaded dendritic cells and antigen-conjugated biodegradable nanoparticles are injected into subjects with T1D at certain doses and concentrations. Thus; i) In individuals with T1D, insulin-producing cells This approach allows for the in vivo programming of T regulatory cells (Tregs), which play a key role in the suppression of autoimmune cells that attack β-cells, to achieve immunotolerance and ultimately prevent autoimmune attacks. Furthermore, it also ensures that an environment that prevents autoimmunity can be created before biochip implantation, ensuring that the tissue engineering product (biochip), which is the basis of the invention, maintains its function in vivo. Decellularized pancreatic extracellular matrix (dpESM) is obtained by pancreatic decellularization to be used as bioink, i.e., printing material (cartridge-loaded material), for 3D bioprinting. Because the technique used in this invention utilizes decellularized material in the scaffolds, it not only offers the possibility of bioprinting with the organ's natural extracellular matrix but also plays a crucial role in the differentiation of mesenchymal stem cells, a component of pancreatic tissue, cultured in this scaffold into beta cells. Extracellular matrix components are generally conserved across species, and even in xenogeneic transplants, they either elicit no immune response or are easily tolerated. To make the matrix obtained after decellularizing the pancreas printable on BB printers, the matrix was lyophilized, powdered, and then converted to a soluble form, giving it gelatinous properties. This form of matrix is highly suitable for supporting cell viability and creating a designable microenvironment. The polymers used in this invention are obtained after organ decellularization and are a unique environment for the organ already present in the body. They are far more than a single polymer like readily available synthetic polymers; they consist of a natural biomaterial with a complex content including many natural polymers such as collagen type 1, collagen type 4, bronectin, glycosaminoglycans, etc. [31]. The chitosan-pDNA nanoparticle ß-cell gene delivery system is being created to enable the differentiation of mesenchymal stem cells in the engineered pancreatic tissue into insulin-secreting ß-cell-like cells. This is a non-viral nanoparticle gene delivery system (genes related to ß-cell differentiation) that is created by using a cationic polymer (chitosan or other non-toxic cationic polymers) instead of a virus. Chitosan-pDNA nanoparticles In the ß-cell gene transfer system, "pDNA" is the short name for plasmid DNA containing beta cell differentiation genes, and pDNA contains Pdx-I and MafA genes or [3 transcription factors involved in cell development. As a result of combining chitosan and pDNAs with ionic gelation, nano-sized Chitosan-Gene Activating Matrix (GAM) as a gene transfer vector consists of dpESM+kit0san-pDNA nanoparticles. The integration of a 3D bioprintable dpESM form with a chitosan-pDNA nanoparticle ß-cell gene delivery system is being achieved to obtain a Gene Activating Matrix (GAM), which stimulates stem cells toward the endocrine pancreas. Simultaneously, the preparation of a bioink by combining the two elements is underway. 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 source cell to generate insulin-secreting cells in the pancreas; in other words, mesenchymal stem cells were differentiated and converted into beta cells by a series of methods. They are also the third element of the bioink. Within the scope of the invention, autologous (taken from the patient himself, taken from the same individual) mesenchymal cells are obtained from the patient's bone marrow or other tissues and directed to differentiate/transform into ß-cells. The properties of MSCs, such as anti-apoptotic, anti-inflammatory, and lack of immunological response in allogeneic transplantations, make these cells Although it is one of the reasons for selection, immunosuppression properties are not the main purpose. Immunosuppression is achieved through in vivo programming of autoimmunity, T regulatory cells (Treg) in a antigen-specific manner in TL. In the invention, a 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+MSC composition); The elements used as bioink, i.e. printing material (material to be loaded into the cartridge), for the 3D bioprinter are integrated in a way that can be printed on the bioprinter and the bioink preparation is completed. A 3D tissue scaffold is produced using bioinks and β-B bioprinters to create a pre-designed 3D tissue piece. These 33 structures, with pre-designed micropatterns and tissue-specific mechanical properties, create a structural environment for mesenchymal stem cells to transform into insulin-secreting pancreatic tissue, providing biocompatible niches within the porous structure. These structures provide important factors such as cell response to the material, adhesion, proliferation on the material, differentiation, and protein synthesis profile. The endocrine pancreas-like functional tissue piece (biochip) is both a novel product created within the scope of the invention and a product used in the treatment of T1D, another aspect of the invention. In this context, it is a biochip that is implanted subcutaneously in a patient with T1D after immunotherapy to be able to secrete insulin in response to glucose and regulate blood sugar to healthy levels. The invention, after reprogramming the immune system that causes autoimmunity in Type 1 diabetes (T1D), is a therapeutic method for the treatment of T1D. It involves the production and transplantation of a functional tissue fragment resembling an endocrine pancreas that can secrete insulin in response to glucose and regulate blood sugar to healthy levels. It also involves the production of glucose-sensitive, insulin-secreting natural tissues by developing next-generation beta cell differentiation techniques in endocrine pancreas engineering. This invention, which targets the treatment of T1D in every aspect, consists of two main sub-studies in detail: immunotherapy and B-cell replacement (replacement of lost tissue). The complete approach to the treatment of TLD, both immunotherapy and beta-cell replacement, is presented. Immunoregulation approaches are reorganized within the scope of the invention and two key points are targeted: 1) In vivo programming of T regulatory cells (Tregs), which play a primary role in immunosuppression, to ensure immunotolerance against autoimmune immune cells (especially autoreactive cytotoxic T cells) that attack insulin-producing ß-cells in individuals with T1D, and ultimately to prevent autoimmune attacks. 2) Creating an environment that prevents autoimmunity before biochip transplantation so that the tissue-engineered biochip (insulin-secreting tissue) can maintain its function in vivo. For beta cell replacement, the development of new generation beta cell differentiation techniques in endocrine pancreas engineering will enable the production and transplantation of a functional endocrine pancreas-like tissue fragment that can regulate blood sugar to healthy levels by secreting insulin in response to glucose. provides. The methodological innovations provided by Özgn in the invention are about creating a tissue engineering product in which more than one technique that has not been tried before is developed and enriched with gene transfer for ß-cell replacement in T1D. Here, a hybrid printing technique is developed by using two separate fields of tissue engineering, decellularization and 3D bioprinting, together, and the production of a bioactive matrix that is rewritten by using stem cells of the pancreas's own extracellular matrix is provided. In the light of all this information, the invention includes the innovations listed below: i. The complete therapeutic approach of the invention for T1D, ii. Production of an endocrine pancreas-like functional tissue part iii. The tissue engineering method used iv. In this tissue transplantation invention, existing autoimmune system cells in individuals with T1D are apoptosis-induced with low-dose radiotherapy, followed by dendritic cell and antigen-conjugated biodegradable nanoparticle-mediated regulatory T cell (Treg) induction, resulting in tolerance of autoreactive T cells in T1D (immune tolerance to ß-cells). This invention overcomes the problems associated with insulin requirement in studies targeting only autoimmunity: a glucose-sensitive insulin-secreting biochip is transplanted into individuals with T1D to compensate for insulin deficiency resulting from the loss of ß-cells that occur with the development of T1D following autoimmunity reprogramming. As a result, a technique is being developed that eliminates the autoimmunity present in T1D, normalizes blood glucose levels, and maintains normal blood glucose levels for the long term. The technique for fabricating an endocrine pancreas-like tissue scaffold for beta-cell differentiation from MSCs is specifically designed for the treatment of T1D. This technique, which allows the solubilization of the extracellular matrix obtained after pancreatic decellularization to be printed in a solution according to pre-designed micropatterns using 3D bioprinters, is the first such method used for the treatment of T1D. Thus, by combining the strengths of both decellularization and 3D bioprinting, a new and powerful hybrid technique for endocrine pancreas tissue engineering is being created. Bioink is printed on 3D bioprinters, and the differentiation stimulation of mesenchymal stem cells in 3D tissue cultured in an endocrine differentiation medium towards the endocrine pancreas is achieved by: i. a gene transfer system for ß-cell differentiation, ii. dpESM, and iii. endocrine medium. In this invention, a cell-free pancreatic extracellular matrix (Decellularized pancreatic extracellular matrix (dpESM)) is obtained by pancreatic decellularization to be used as bioink, i.e., printing material (material to be loaded into the cartridge), for the 3D bioprinter. Because decellularized material is used in tissue scaffolds produced using this technique, it not only offers the possibility of bioprinting using the organ's natural extracellular matrix, but also plays a crucial role in the differentiation of mesenchymal stem cells, a component of the tissue cultured and formed within the scaffold, into beta cells. Extracellular matrix components are generally conserved across species and, even in xenogeneic transplants, either elicit no immune response or are easily tolerated. After dpESM is obtained, the powdered product is lyophilized and then turned into a printing solution to obtain a bioink form that optimizes cell viability and printability of the material on 313 bioprinters. This allows for a homogeneous composition in the tissue produced while achieving a form suitable for matrix printing. Following immunotherapy, a functional endocrine pancreas-like tissue fragment (biochip) is obtained via subcutaneous transplantation to enable the T1D patient to secrete insulin in response to glucose and maintain healthy blood sugar levels. Biochips are biocompatible and living systems, free of plastic or metal-like materials, designed for implantation into the living body, and are transportable biological systems. The biochip produced within the scope of the invention aims to provide beta cell replacement and eliminate the autoimmune response through the interactions of nanoparticles and dendritic cells and T regulatory cells. The invention does not approach T1D from a single perspective, but rather views the disease as a whole and addresses it in a complex and multi-stage manner. The tissue scaffold used within the scope of the invention is produced using BB bioprinters according to the desired micropatterns after the matrix obtained by decellularization of the organ is pulverized (the similarity between the aforementioned patent and our patent in terms of matrix goes only so far). It is designed electronically using software and produced using BB bioprinters. This integrated method allows for the integration of a non-viral gene delivery system into the matrix. Using this system brings many advantages: i. The solution obtained by solubilizing the decellularized pancreatic matrix constitutes the base material of the tissue scaffold. ii. The ability to redesign the matrix in a computer environment allows for the final shape of the produced chips, allowing for the design of the matrix according to the region to be transferred to the living organism. iii. The matrix content allows the addition of various polymers, growth factors, and media stabilizers, if necessary, to the solution obtained by solubilizing the decellularized pancreatic matrix. In other words, the system used is suitable for enriching the medium. iv. Following matrix recellularization, the method employed within the scope of the invention also enables the integration of a gene delivery system into the decellularized pancreatic extracellular matrix solution, which will facilitate the transformation of seeded cells into insulin-producing beta cells. Furthermore, the gene delivery system is preferred as a non-viral system for the production of clinically suitable biochips. Briefly, the advantages and innovations provided by the invention are: Many studies on F TlD have attempted allogeneic beta cell, islet, or pancreas transplantation, and because such transplants are allogeneic, they often cause severe immune responses. Patients are forced to take immunosuppressive medications. The invention utilizes autologous mesenchymal stem cells taken from patients and then transformed into beta cells. Therefore, the tissues produced are personalized and do not trigger an immune response when transplanted into the patient. Individuals with TLD are forced to use exogenously administered insulin throughout their lives. Thanks to the tissue produced, the body's own insulin needs will be produced according to blood glucose levels, and patients will be able to continue their lives without the need for exogenous insulin. Considering that the amount of insulin produced is quite high, it creates a significant burden on our country's economy. Thanks to the tissue produced, this economic loss is also prevented. Beta cells differentiated from BF MSCs are cultivated in SB environments, unlike 2D culture methods. This offers the opportunity to better mimic the body, unlike many previous studies. I› The invention is a technique specifically designed for the treatment of TLD. The insulin-secreting endocrine pancreas-like scaffold production technique, which will be used for beta cell differentiation from MSCs. The solution produced by solubilizing the extracellular matrix obtained after decellularization of the pancreas is cultured in 3D cultures. This technique, which allows for the printing of pre-designed micropatterns in bioprinters, is the first method used for the treatment of T1D. Thus, by combining the strengths of both decellularization and 3D bioprinting, a new and powerful hybrid technique for endocrine pancreatic tissue engineering is being created. Gene-activating matrices (GAýM: dpESM + chitosan-pDNA nanoparticles) are obtained by augmenting the natural (non-synthetic) polymer structure of the 3D bioprinted pancreas-derived matrix (dpESM) with non-viral gene delivery systems (chitosan pDNA nanoparticles). The tissue scaffold printed from this matrix provides a unique microenvironment for beta cell differentiation from mesenchymal stem cells, thus developing a new generation of beta cell differentiation method. 3D bioprinting; tissue scaffold The method's significant advantages include the ability to place cells in the printed tissue during the printing phase, the ability to adjust the scaffold to the desired composition, and the ability to design it according to the desired micropatterns. It provides much more biocompatible niches than matrices created using other methods. However, the material used and the composition created in 3D bioprinting are crucial. Because decellularized (non-synthetic, natural) material is used in the scaffolds produced with this technique, it is crucial that it offers the possibility of bioprinting with the organ's natural extracellular matrix. This is because the ratios of extracellular matrix proteins (such as collagen, fibronectin, laminin, etc.) in the tissue are preserved, and thanks to the cells contained in the bioink, every region of the tissue can be easily cellularized. Extracellular matrix components are generally conserved across species and can be replicated even in xenogeneic transplants. They either elicit no immune response or are easily tolerated. This technique allows the use of decellularized extracellular matrices derived from different species while preserving the natural structure. This overcomes the problems encountered in allogeneic and even xenogeneic organ/tissue transplants. The invention is also a cellular treatment product for T1D7. Endocrine pancreas engineering involves the production of a functional endocrine pancreas-like tissue fragment (biochip) capable of regulating blood sugar to healthy levels by secreting insulin in response to glucose, using next-generation beta cell differentiation techniques. This "product", created by the development of many techniques, constitutes a new concept in itself. The problems related to the insulin requirement in studies targeting only autoimmunity are overcome in this invention as follows: The invention presents a complete approach to the treatment of T1D7, both as immunotherapy and beta-cell replacement. 1) Preventing autoimmune attacks by in vivo programming of T regulatory cells (Treg) in individuals developing T1D, 2) Creating an environment that prevents autoimmunity before biochip transplantation so that the tissue-engineered biochip (insulin-secreting tissue), which is the basis of the invention, can maintain its function in vivo, 3) Developing new generation beta cell differentiation techniques in endocrine pancreas engineering, which can secrete insulin in response to glucose and regulate blood sugar to a healthy level. The invention enables the production and transplantation of functional endocrine pancreas-like tissue parts. In the invention, ß-cell differentiation from MSCs is achieved on a tissue scaffold that provides a microenvironment very close to the natural one, while dendritic cell- and/or antigen-conjugated biodegradable nanoparticle-mediated Tregderin re-regulation is provided to achieve immunotolerance of autoreactive immune cells against beta cells in T1D. Therefore, the invention can target the treatment of T1D disease as a whole. Thus, a new strategy is provided that enables the treatment of T1D disease by generating personalized tissues and achieving tolerance of autoimmune T cells. Application of the Invention in Industry Since the product produced within the scope of the invention is a living tissue that can be produced specifically for the patient and can secrete insulin, the expenses for commercially sold insulin are significantly reduced. It has the potential to prevent a significant proportion of diabetes. In addition, since other complications seen in chronic periods in diabetic patients can be prevented, the expenses incurred for the treatment of these diseases will be reduced or even 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 method suitable for biofabrication. For each patient, personalized tissues can be produced by replacing only the patient's cells. If the clinical use targets 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 also prevent many secondary complications that arise in the chronic period before they develop. Thus, it will bring a high added value to our country. The target audience of the invention is TLD patients, but although the invention is not aimed at the treatment of TLD disease, Although it is a product that has been developed, it will also serve as a model for the treatment of many other autoimmune diseases. The invention is the product of a highly complex, multi-step approach that combines both beta cell replacement and immunotherapy, two important approaches developed to date, with current tissue engineering techniques. It also reorganizes existing tissue engineering techniques to make them suitable for clinical use, resulting in a brand new, multi-step approach. It is a multidisciplinary study that can be cited as an example of studies in the field of stem cells, immunotherapy, tissue engineering, and TlD.