The Human Pancreas as a Source of Protolerogenic Extracellular Matrix Scaffold for a New-generation Bioartificial Endocrine Pancreas

Abstract

OBJECTIVE Our study aims at producing acellular extracellular matrix scaffolds from the human pancreas (hpaECMs), as a first critical step towards the production of a new generation, fully human-derived bio-artificial endocrine pancreas (BAEP). In this BAEP, the hardware will be represented by hpaECMs, while the software will consist in the cellular compartment generated from patient’s own cells. SUMMARY BACKGROUND DATA ECM-based scaffolds obtained through the decellularization of native organs have become the favored platform in the field of complex organ bioengineering. However, the paradigm is now switching from the porcine to the human model. METHODS To achieve our goal, human pancreata were decellularized with Triton-based solution and thoroughly characterized. Primary endpoints were: complete cell and DNA clearance, preservation of ECM components, growth factors (GFs) and stiffness, ability to induce angiogenesis, conservation of the framework of the innate vasculature, and immunogenicity. Secondary endpoint was hpaECMs’ ability to sustain growth and function of human islet and human primary pancreatic endothelial cells (hPPEC). RESULTS Results show that hpaECMs can be successfully and consistently produced from human pancreata, maintain their innate molecular and spatial framework and stiffness, as well as vital GFs. Importantly, hpaECMs inhibit human naïve CD4+ T cell expansion in response to polyclonal stimuli by inducing their apoptosis and promoting their conversion into regulatory T cells. hpaECMs are cytocompatible and supportive of representative pancreatic cell types. DISCUSSION We therefore conclude that hpaECMs has the potential to become an ideal platform for investigations aiming at the manufacturing of a regenerative medicine-inspired BAEP.

Strategies based on organ decellularization and recellularization

“…Furthermore, they seeded islets on scaffold cubes and observed insulin secretion after stimulation with high glucose concentrations. The authors also showed a reendothelialization with human pancreatic endothelial cells in vitro over a culture period of 6 days .…”

“…Only a few publications have examined the decellularization of whole pancreas and the subsequent repopulation with islets of Langerhans. Protocols for perfusion‐based decellularization of the mouse, rat, pig and human pancreas have been published , but use different detergents and times to decellularize the pancreas. Detergents include nonionic tensides such as Triton X‐100 and enzymatic agents such as DNAse.…”

Heterogeneity of Scaffold Biomaterials in Tissue Engineering

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Figure A2Immune properties of ECM scaffolds obtained from the human pancreas. ( A ) Representative plots and ( B ) quantitation of carboxyfluorescein succinimidyl ester (CFSE) dilution in CFSE-labeled human naïve CD4 + T cells stimulated with anti-CD3/anti-CD28-coated beads cultured with or without pulverized ECM; ( C ) Representative plots and ( D ) quantitation of Annexin-V and 7-AAD expression as a measure of apoptosis/necrosis in human naïve CD4 + T cells stimulated with anti-CD3/anti-CD28-coated beads cultured with or without ECM; ( E ) Representative plots and ( F ) quantitation of FoxP3 expression in CD4 + CD25 + cells converted from human naïve CD4 + T cells stimulated with anti-CD3/anti-CD28-coated beads and IL2 cultured with or without ECM (with permission from Peloso et al Ann Surg 2015, ahead of print) [145]. * denotes p < 0.05.

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Bioengineering the Pancreas: Cell-on-Scaffold Technology

“…In 2015, our group has explored the use of discarded pancreas as a source from which obtain acellular whole organ scaffolds [130]. Compared to data achieved on the porcine model, an additional inlet access was demonstrated crucial switching from a two-way perfusion (pancreatic duct and superior mesenteric vein) to a three-way perfusion system (pancreatic duct, superior mesenteric artery and splenic artery).…”