Speaker
Description
Pancreatic islet transplantation holds significant potential for treating insulin-dependent diabetes, but its clinical utility remains limited by donor scarcity, immune rejection, and the need for lifelong immunosuppression. To overcome these challenges, stem cell-derived islets, particularly those differentiated from human induced pluripotent stem cells (iPSCs) engineered for hypoimmunogenicity through CRISPR-Cas9-mediated gene editing, have emerged as a promising alternative source. However, despite their immune-evasive properties, SC-derived islets often exhibit incomplete maturation compared to primary islets, highlighting the need for additional engineering strategies to ensure functional efficacy in vivo. Here, we present a bioengineered islet platform termed hypoimmune iPSC-derived bioprinted islet-like cellular aggregates (HIBICA), which integrates gene-edited hypoimmune iPSC-derived islets with a tissue-specific biochemical niche and a scalable fabrication strategy to generate transplant-ready islet constructs. To promote functional maturation in vitro, we employed pancreatic tissue-derived extracellular matrix (pdECM) as a physiologically relevant microenvironment. Although human-derived pdECM effectively supports insulin-secreting-β cell identity, its limited availability poses challenges for translational scalability. Therefore, we evaluated porcine-derived pdECM as an alternative, and through comparative proteomic profiling and gene ontology analysis, confirmed its compositional similarity to the human-derived pdECM. Notably, porcine-derived pdECM demonstrated significant glucoregulatory support compared to collagen type I, providing a functionally enriched matrix that effectively facilitated the maturation of hypoimmune iPSC-derived islets to levels comparable with those supported by human-derived pdECM. To address limitations in conventional islet aggregate formation methods, such as suspension culture utilizing orbital shakers or spinner flasks, which require at least 24 hours for aggregate formation and often result in heterogeneous aggregate size and unintended clustering that can lead to hypoxic conditions within larger aggregates, we applied embedded 3D bioprinting to rapidly and precisely generate islet-like aggregates within the pdECM bioink. This approach enabled high-throughput fabrication of uniformly sized islet-like aggregates, achieving the production of over one aggregate per second with consistent architecture. The embedded HIBICA maintain their endocrine phenotype, showing stable expression of key hormonal markers (e.g., insulin, glucagon, and somatostatin) along with sustained expression of immune checkpoint markers. Following transplantation into NSG mice, HIBICA outperformed both non-edited SC-islet grafts and hypoimmune islets transplanted without matrix support, exhibiting sustained in vivo insulin secretion and improved graft stability. These findings demonstrate the synergistic effect of immune engineering and biochemical niche optimization coupled with geometrical guidance in enabling long-term graft function without immunosuppression. In conclusion, HIBICA offers a comprehensive platform for advancing SC-derived islet transplantation by combining hypoimmune genetic modifications, tissue-specific matrix support, and scalable fabrication. Furthermore, this platform opens opportunities for allogeneic, off-the-shelf islet therapies, and may serve as a foundation for future combinatorial approaches involving hypoimmune vascularization and integration with microencapsulation systems, expanding therapeutic flexibility in regenerative diabetes care.
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