Over the project, partners developed and refined each component of the bioartificial pancreas. Early work established pipelines for human amniotic epithelial and mesenchymal stem cells and generated immunomodulatory variants with enhanced HLA-G and PD-L1. In vitro assays showed both native and modified hAMSCs supported β-cell viability and insulin secretion, though with only modest protection against cytotoxic T lymphocytes. Given these results and the regulatory and manufacturing complexity of a third-party allogeneic source, the partners excluded amniotic cells. This ensured the therapeutic effect was attributable to β-cells, endothelial support and Amniogel, while keeping the product clinically feasible and translatable.
Due to streamlined design, organoid generation advanced, markedly improving insulin secretion. Porcine organoids from neonatal porcine islet cells and blood outgrowth endothelial cells (BOECs) reversed diabetes in mice faster than free neonatal islets, demonstrating the advantage of vascularised constructs.
A GMP-compatible process for producing Amniogel established. Batches showed reproducible composition, retaining extracellular matrix proteins, tunable stiffness and pore size, and stable support. Functionally, Amniogel restored cell–matrix interactions, upregulated β1-integrin, reduced apoptosis, and improved insulin secretion. It enhanced β-cell–β-cell and β-cell–endothelial communication, supported by BOEC-driven laminin deposition. In vitro assays also showed Amniogel delayed CD8⁺ T-cell migration and reduced cytotoxicity toward mismatched β-cells, validating its role as both structural and immunological barrier.
These properties enabled vascularised endocrine constructs of intact islets and BOECs in Amniogel. A human BAP was generated with native islets and BOECs, while a porcine BAP combined neonatal porcine islets with BOECs. When transplanted subcutaneously in diabetic mice, both restored normoglycaemia, confirming the importance of ECM support and pre-established vascularisation for extrahepatic sites. Safety studies showed Amniogel supports rodent, human, and porcine islets with excellent biocompatibility. Immune-evasion studies revealed MHC-I down-regulation reduces CD8⁺ T-cell graft destruction without provoking NK rejection, and Amniogel encapsulation can delay, though not fully prevent, graft rejection in humanised mouse models.
The project generated strong dissemination outputs. Results were published in peer-reviewed journals and presented internationally, raising visibility in transplantation, regenerative medicine, and biomaterials. Ethical and legal analyses produced publications and a forthcoming guidance document on responsible development of bioartificial organs. A regulatory roadmap was drafted for BAP classification as an ATMP, and spheroid scalability was demonstrated with the GMP-compliant Sphericalplate 5D platform.
Stakeholder engagement included patient events such as World Diabetes Days, the project website with a patient area, a policy brief, and participation in European Researchers’ Night. These ensured broad dissemination, integrated patient and policymaker feedback, and positioned VANGUARD as a leading contributor to the evolving ATMP landscape.
