Stem cell-based therapies for type 1 diabetes: Progress in differentiation, clinical translation, and immune protection.

Currently it is less advanced to understand the pathology of lifestyle disease by using iPS cells because there is partly less direct connection between life style disease and iPS cells. So much more scientists focus on regenerative medicine such as beta cells therapy using iPS cells technologies. It will be indeed a powerful tool to generate beta cells from iPS cells as even in type2 diabetes patients, hyposecretion of insulin from beta cells in pancreas is one of causes. Another reason is complexity of the pathology of life style disease. There are a lot of reasons to cause lifestyle disease. Lifestyle diseases include cancer, chronic liver disease, Type 2 diabetes, heart disease, metabolic syndrome, chronic renal failure, stroke, and obesity. Since obesity is one of major causes of lifestyle diseases, we want to focus on adipogenesis from iPS cells in this review. We analysed and established the differentiation protocol into adipocytes from mouse ES cells and human iPS cells. The other point in this review is the starting pluripotent cells for differentiation. Quality of pluripotent stem cells are one of most critical factors to succeed in getting well-differentiated cells. Recently, we have developed new naive human pluripotent stem cells (PSC),"Reset cells". Naive PSC have more similar to human epibast cells than conventional human PSC. They will be more ideal cells for differentiation because of their hypomethylated status and earlier stage of development.

Recent trends and advances in type 1 diabetes therapeutics: A comprehensive review

Differentiation of pluripotent stem cells (PSCs) into β cells could provide insulin independence for type 1 diabetes (T1D) patients. This approach would reduce the clinical complications that most patients managed on intensive insulin therapy (IIT) face. However, bottlenecks of PSC manufacturing and limited engraftment of encapsulated cells hinder the long-term effectiveness of these therapies. A bioprocess decision-support tool is combined with a disease state-transition model to evaluate the cost-effectiveness of the stem cell-based therapy against IIT. Clinical effectiveness is assessed in quality-adjusted life years (QALYs). Manufacturing costs per patient reduce from $430 000 to $160 000 with optimization of batch size and annual demand. For 96% of the patients, cell therapy improves the quality of life compared to IIT. Cost savings are achieved for 2% of the population through prevention of renal disease. The therapy is cost-effective for 3.4% of patients when a willingness to pay (WTP) of up to $150 000 per QALY is considered. A 75% cost reduction in the cell therapy price increases cost-effectiveness likelihood to 51% at $100 000 per QALY. This study highlights the need for scalable manufacturing platforms for stem cell therapies, as well as to prioritizing access to the therapy to patients with an increased likelihood of costly complications.

Human pluripotent stem cells represent a potential source of pancreatic beta cells for the treatment of type 1 diabetes. Methods for generating pancreatic endoderm from pluripotent stem cells are becoming increasingly robust, although the general applicability of any given method to a broad range of stem cell lines remains a challenge. Therefore, a deeper understanding of the molecular mechanisms controlling the commitment of cells to pancreatic endoderm is desirable. A key step in this process is the co-ordinated up-regulation of two transcription factors, PDX1 and NKX6.1; factors that together identify cells fated to become pancreatic endoderm and to later give rise to endocrine cells. In this thesis, we examined the effects of overexpressing PDX1 and NKX6.1 during the course of PSC differentiation. We employed the use of a number of transgenic systems, namely the doxycycline inducible Tet-On expression system, Destabilisation Domains (DD) and Estrogen Receptor (ER) fusion proteins. Our experiments demonstrated a potential for each of the 3 expression systems, however we encountered drawbacks within each one preventing us from establishing a definitive conclusion as to the affects of over-expressing PDX1 and NKX6.1 during PSC differentiation. We conclude that only once both the expression systems and differentiation protocols are optimised will it be possible to determine the effects of co-expressing these two factors during the course of pancreatic differentiation.

The pancreas plays a central role in metabolism. The exocrine pancreas, composed of ductal and acinar cells, secretes and delivers digestive enzymes into the duodenum where they contribute to food digestion. The endocrine pancreas, constituted by the islets of Langerhans, secretes hormones in the blood to control glucose levels. In particular, insulin is the hormone produced by β-cells that instructs the peripheral tissues to uptake circulating glucose. Diabetes mellitus is a heterogeneous disease characterized by deregulated glucose homeostasis due to an impaired insulin function. The advancements in clinical practice have made of diabetes a chronic, instead of a lethal, disease; yet no definitive cure is available. Beta-cell transplantation offers promising results, especially for type 1 diabetes where β-cells are selectively eliminated by an autoimmune attack. Unfortunately its application suffers from the scarcity of transplantable pancreas/islets from cadaveric donors. Human embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) would constitute unlimited sources of transplantable β-cells, but the available differentiation protocols are not yet optimal, especially with regards to the numbers and the functionality of the generated β-cells. It is anticipated that a stepwise protocol would succeed in generating mature β-cells in vitro if it is capable of fully mimicking the embryonic development of these cells. Many aspects of pancreatic organogenesis have been elucidated and could serve as a guide for driving the commitment to β-cells. Unfortunately, pancreatic progenitors in vitro do not behave as in the intricate context of an embryo. This severely limits both the understanding of pancreatic development at the single-cell level and our ability in manipulating the process. The aim of our work was to develop in vitro methodologies to sustain pancreatic progenitor culture and expansion and to establish conditions to manipulate their progeny. To this aim, we combined an informed approach based on developmental biology and an empirical one. We first observed that pancreatic epithelial explants from embryonic day 10.5 mice could be cultured in presence of Matrigel™ and exogenous FGFs even in the absence of the mesenchyme that normally surrounds the epithelium. Notably, the removal of mesenchyme did not affect the endocrine commitment pattern of pancreatic progenitors. We then defined culture conditions allowing for the expansion of dissociated embryonic pancreatic progenitors in a three-dimensional Matrigel™-based environment. Under these conditions, progenitors proliferated and self-organized to generate pancreatic organoids containing both progenitors and differentiated cells, mostly exocrine, after 7 days of culture. When grafted into recipient pancreatic explants, the cultured progenitors contributed to the three epithelial pancreatic lineages (ductal, endocrine, acinar), integrating seamlessly into the endogenous cellular network. Thus, cultured progenitors retained their potential to differentiate into endocrine cells, but expressed it only in the appropriate niche. Subsequent removal of single components from the culture system led to the identification of some essential requirements for the maintenance/expansion of dissociated pancreatic progenitors, such as a strong FGF signaling and the Rho-associated kinase (ROCK) inhibitor Y-27632. In addition to this, we observed that only small clusters of pancreatic progenitors, but not single cells, were able to generate pancreatic organoids, suggesting a pivotal role for intercellular signaling in progenitor maintenance and expansion. By manipulating the components of the medium, we could affect fate commitment. In particular, the removal of FGF1 increased the yield of endocrine cells generated in vitro. In our search for a full control over the in vitro niche, we explored chemically defined matrices to replace Matrigel™. We showed that pancreatic progenitors could be cultured on laminin-functionalized hydrogels, although less efficiently than in Matrigel™. Moreover, differentiation into exocrine and endocrine cells occurred spontaneously under these conditions. By comparing Matrigel™, hydrogel microwells and two-dimensional culture systems, we uncovered the importance of a tridimensional architecture for pancreatic progenitor maintenance. We showed that pancreatic progenitors lost their identity as they flattened onto 2D surfaces, whereas 3D culture systems maintained the epithelial character of pancreatic progenitors and allowed the acquisition of apical polarization. To conclude, our work provides the first detailed characterization of in vitro culture systems for expansion and manipulation of dissociated embryonic pancreatic progenitors. Further implementation will hopefully allow the establishment of a fully-defined in vitro niche for developing pancreas with potential fundamental implications for the expansion of ES-derived pancreatic progenitors and their differentiation into functional β-cells.

Introduction: Type 1 diabetes (T1D) is a disease characterized by elevated blood glucose due to insufficient insulin release from pancreatic β-cells. Transplantation of cadaveric islets demonstrates that cell therapy can fully reverse hyperglycemia. However, limited cell supply, immune rejection of implanted allogeneic cells, and cell survival represent major challenges. Cell encapsulation has great potential to overcome these challenges by blocking immune cell access to the grafts while allowing nutrient exchange and secreted products from the implanted cells to be delivered to the body. In this study, we use a unique microfluidic bioprinting technology to precisely control the placement of cells and biomaterials within 3D tissues with micro-architectures optimized for cellular fitness and immune protection. Methods: Living tissues consisting of fibres with a cell-containing core and immune-protective alginate-based shell were generated using Aspect Biosystems’ RX1 bioprinter technology. Core-shell fibres with reaggregated primary human pancreatic islets or embryonic stem cell-derived β-cells were tested in vitro using viability and functional (glucose-stimulated insulin secretion, GSIS) assays. Bioprinted tissues were then implanted into the IP space or omentum of streptozotocin (STZ)-induced diabetic mice and rats, respectively. Glucose homeostasis, body weight, and human C-peptide secretion were monitored for up to 3 months following implantion. Retrieved grafts were fixed and analyzed by histology (H&E, Masson’s trichrome stain) and immunohistochemistry (α-SMA, CD45) to quantify fibrotic encapsulation and immune cell infiltration. Results: Bioprinted tissues supported viability and dynamic insulin secretion of cells in vitro up to 28 days. When transplanted into immunodeficient and immunocompetent diabetic rodents, bioprinted tissues containing reaggregated human islets successfully regulated blood glucose for up to 3 months (Figure 1), although normoglycemia was only sustained in a subset of immunocompetent animals and associated with variable fibrosis. Post-retrieval viability stain, assessment of GSIS, and histology revealed high viability and functionality of implanted cells, and the absence of leukocyte infiltration through the shell. Discussion: This is the first study showing a fully 3D bioprinted tissue composed of a core/shell fibre can successfully deliver a therapeutic dose of xenogeneic cells into a diabetic animal (Figure 2). The unique features of the microfluidic technology were leveraged to bioprint an implantable and retrievable tissue patch that merges the benefits of a single fibre (cell fitness, access to nutrients, immune-protection, perm-selectivity) with those provided by a 3D structure (retrievability, ease of implant, structural integrity). The promising results obtained in rodent studies warrant further investigation to minimize fibrosis and explore the scaling-up of bioprinted tissues in large animal models of T1D.NRC-IRAP. NSERC. Stem Cell Network. Genome BC.

ground: Type 1 diabetes is the result of an autoimmune attack against the insulin-producing beta cells of the pancreas. Current treatment for patients with type 1 diabetes typically involves a rigorous and invasive regimen of testing blood glucose levels many times a day along with injections of recombinant insulin. Many recent researches have shown that stem cell therapy can be the best choice for treatment of this disease. The aims of this research were investigating regeneration of pancreatic beta cells of type 1 diabetic rabbits after stem cell transplantation. Materials and Methods: 32 rabbits weighting an average of (2.5 - 3 kg) were used in this experimental study, and divided into 2 groups as follows; group A ( contains 16 controlled diabetic rabbits received insulin as a treatment ) and group B ( contains 16 diabetic rabbits received autologous mesenchymal stem cells as a treatment).The induction of diabetes was achieved by a single dose of intravenous injection of the Alloxan, which was administered to the rabbits via the marginal ear vein, mesenchymal stem cells were differentiated into insulin - producing cells and reimplanted into the rabbits of group B with daily monitoring of blood glucose level and body weight. Results: The insulin - producing cells regulated the hyperglycemia resulted from diabetic rabbits , 7 to 9 days after reimplantation the blood glucose level were decreased from about( 400 mg/dl into 180 mg/dl). Conclusions: Islet-like functional cells can be differentiate d from bone-marrow mesenchymal stem cells (MSCs), which may be a new procedure for clinical diabetes stem -cell therapy, these cells controlled blood glucose level in diabetic rabbits as the effect of insulin. MSCs play an important role in diabetes therapy by islet differentiation and

The identification of human embryonic stem cells and reprogramming technology to obtain induced pluripotent stem cells from adult somatic cells have provided unique opportunity to create human disease models, gene editing strategies and cell therapy options.Development of pluripotent stem cells from somatic cells and genomic manipulation tools enabled to use site specific nucleases in the cell therapy research. Identification of efficient gene manipulation, safe differentiation and use will provide a novel strategy to treat many diseases in the near future. Current available registered clinical trials clearly indicate the need for pluripotent stem cell and gene therapy treatment options. Although gene editing based pluripotent stem cell research is a popular field for research worldwide, improvement of clinical approaches for treatment still remains to be investigated. In this review, we summarized the current situation of gene editing based pluripotent cell therapy developments and applications in clinics.

BackgroundType 1 diabetes (T1D) is a chronic autoimmune disorder resulting in the destruction of pancreatic beta cells, leading to absolute insulin deficiency. Despite advances in exogenous insulin therapy, patients continue to face significant challenges, including glycemic variability, risk of hypoglycemia, and long-term complications. The complex interplay between genetic, immunological, and environmental factors in T1D pathogenesis underscores the need for more targeted and durable therapeutic strategies.PurposeThis review aims to critically evaluate recent breakthroughs in cell-based, immunomodulatory, and gene therapy approaches for T1D, moving beyond insulin-centric management. It seeks to analyze the mechanisms, benefits, limitations, and translational potential of these emerging modalities, alongside advances in insulin formulation and delivery systems.Main bodyInnovations in cell-based therapies, notably stem cell-derived beta cell replacement and porcine islet xenotransplantation, are advancing toward clinical translation, supported by encapsulation technologies that enhance cell viability and immune protection. Parallel progress in immunomodulatory approaches includes both antigen-specific (oral insulin, GAD65, and proinsulin peptide therapies) and non-antigen-specific interventions (anti-CD3 monoclonal antibodies, engineered regulatory T cells). Combination immunotherapies and the advent of Teplizumab have demonstrated potential for delaying disease progression and preserving beta cell function. Gene therapy and genome editing (CRISPR/Cas9) are being refined for beta cell regeneration, protection, and immune tolerance induction, though challenges of safety, immunogenicity, and off-target effects persist. Furthermore, advances in glucose-responsive “smart” insulins, ultra-rapid-acting formulations, and automated delivery systems are optimizing glycemic control. Emerging fields such as nanotechnology, bioartificial pancreas development, and microbiome modulation further expand the T1D therapeutic landscape.ConclusionThe integration of cell-based, genetic, and immunological therapies holds promise to fundamentally alter T1D management, offering prospects for long-term remission or cure. However, translational hurdles—especially immunological, ethical, and regulatory concerns—must be addressed. Multimodal, patient-tailored strategies and robust clinical validation are critical for future progress.Graphical

CRISPR/Cas9-Mediated Genome Editing Corrects Dystrophin Mutation in Skeletal Muscle Stem Cells in a Mouse Model of Muscle Dystrophy.

<p>Type 1 Diabetes (T1D) is an autoimmune disorder in which the pancreatic β-cells are destroyed by the body's immune system. The reduced number of β-cells leads to inadequate insulin secretion and high glucose levels in the body. The requirement of insulin injection throughout life and lack of donors for islet transplantations has prompted a search for more accessible and available sources of insulin producing cells that can be transplanted in T1D patients. To that end, the discovery of induced pluripotent stem (iPS) cells has provided a potential source of precursors for cell therapy for T1D. iPS cells are reprogrammed somatic cells which can be transplanted back into the patient from whom the somatic cells were initially derived, thus potentially avoiding immune rejection when transplanted. As a potential therapy for T1D, we aim to derive insulin producing cells (IPCs) from human iPS cells. In contrast to the conventional two dimensional (2D) cell culture systems used in many iPS derived IPC studies, the inner cell mass (ICM) from which various organs differentiate during embryogenesis is a cluster of cells that enables signaling crosstalk between cells of different types. Three dimensional (3D) cell culture systems allows cells to form cell clusters that promote cell - cell signaling. Hence, we hypothesized that 3D cell culture systems will yield better efficiency of differentiation to functional IPCs <em>in vitro</em> than 2D cultures.</p><p>Initially, the synthetic polymers sodium alginate and matrigel were analyzed for their ability to enable cell clustering to establish 3D cell culture systems. The 3D cell environment established using matrigel was used for the differentiation of human iPS cells to Insulin Producing Cells (IPC). The cells were first converted to endodermal cells. A mixture of growth factors then induced the differentiation of endodermal cells to pancreatic cells. The pancreatic cells were converted to IPCs that resemble pancreatic β-cells. Our 3D differentiated IPCs strongly expressed pancreatic endocrine transcription factors and pancreatic hormones. The IPCs also produced insulin when exposed to a high glucose environment. But the number of IPCs obtained at the end of the differentiation was low.</p><p>Hence, our results demonstrate that 3D differentiation generates functional IPCs <em>in vitro</em> unlike 2D differentiation. In the future we aim to improve the percentage of IPCs that we generate from the 3D differentiation. Our expectation is that these cells will be able to cure hyperglycemia in diabetic mice more rapidly compared to the 2D differentiated cells owing to their proven insulin production in the presence of a high glucose environment in vitro.</p>

Purification of human iPSC-derived cells at large scale using microRNA switch and magnetic-activated cell sorting.

Rapid and Efficient Generation of Functional Motor Neurons From Human Pluripotent Stem Cells Using Gene Delivered Transcription Factor Codes

Type 1 diabetes (T1D) is a chronic autoimmune disease affecting millions of people worldwide. The condition results from the immune-mediated destruction of pancreatic β-cells, leading to insulin deficiency and lifelong dependency on insulin therapy. Despite advances in insulin delivery and glucose monitoring technologies, many patients struggle to maintain optimal glycemic control, and severe hypoglycemia remains a persistent risk. Islet transplantation offers a potential alternative but is limited by donor shortages and the need for lifelong immunosuppression. Pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), offer a renewable and scalable source of insulin-producing β-cells. These cells can be differentiated in vitro and have demonstrated glucose-responsive insulin secretion and the ability to reverse diabetes in animal models. Recent years have seen the translation of PSC-derived β-cell therapies into clinical trials, with several promising candidates including VX-880, VC-02, CTX-211 and OZTx-410 under investigation. These approaches include both allogeneic and autologous strategies as well as gene-edited and encapsulated cell delivery systems designed to enhance cell survival and minimize immune rejection. This article reviews the latest advances in PSC-based cell therapies for T1D with a focus on differentiation protocols, preclinical studies and ongoing clinical trials. It also discusses current challenges including immune protection, vascular integration, and scalability and outlines future directions for achieving a functional cure through regenerative medicine.

WNT3 Is a Biomarker Capable of Predicting the Definitive Endoderm Differentiation Potential of hESCs