Decellularized pancreas as a native extracellular matrix scaffold for pancreatic islet seeding and culture

Abstract

Diabetes mellitus involves the loss of function and/or absolute numbers of insulin-producing β cells in pancreatic islets. Islet transplantation is currently being investigated as a potential cure, and advances in tissue engineering methods can be used to improve pancreatic islets survival and functionality. Transplanted islets experience anoikis, hypoxia, and inflammation-mediated immune response, leading to early damage and subsequent failure of the graft. Recent development in tissue engineering enables the use of decellularized organs as scaffolds for cell therapies. Decellularized pancreas could be a suitable scaffold as it can retain the native extracellular matrix and vasculature. In this study, mouse pancreata were decellularized by perfusion using 0.5% sodium dodecyl sulfate. Different characterizations revealed that the resulting matrix was free of cells and retained part of the pancreas extracellular matrix including the vasculature and its internal elastic basal lamina, the ducts with their basal membrane, and the glycosaminoglycan and collagen structures. Islets were infused into the ductal system of decellularized pancreata, and glucose-stimulated insulin secretion results confirmed their functionality after 48 hr. Also, recellularizing the decellularized pancreas with green fluorescent protein-tagged INS-1 cells and culturing the system over 120 days confirmed the biocompatibility and non-toxic nature of the scaffold. Green fluorescent protein-tagged INS-1 cells formed pseudoislets that were, over time, budding out of the decellularized pancreata. Decellularized pancreatic scaffolds seeded with endocrine pancreatic tissue could be a potential bioengineered organ for transplantation.

Strategies based on organ decellularization and recellularization

“…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.…”

“…Similarly, only a few publications have examined the decellularization and repopulation of the whole pancreas with islets of Langerhans . Napierala et al .…”

Using GRGDSPC peptides to improve re‐endothelialization of decellularized pancreatic scaffolds

“…Over the past decade, the fields of regenerative medicine and tissue engineering have offered new strategies for the generation of an insulin-secreting organ. 7,8 The creation of a three-dimensional decellularized pancreatic bioscaffold is a new approach that has shown promising success in recent years. 9 Natural tissue extracellular matrices (ECMs) in the decellularized scaffold possess a preexisting architectural structure with organ-specific anatomical and biochemical properties.…”

“…Clearly, replacement with functional pancreatic tissue should be proposed as a regenerative therapy that could restore the physiological regulation of the blood sugar level. Over the past decade, the fields of regenerative medicine and tissue engineering have offered new strategies for the generation of an insulin‐secreting organ …”

Tissue and organ decellularization in regenerative medicine

“…[35][36][37][38] Recently published work from our group demonstrated that, islets infused into the ductal system of decellularized pancreata were functional by releasing insulin in response to glucose stimulation after 48 days in vitro. 39…”