Engineering an endocrine Neo-Pancreas by repopulation of a decellularized rat pancreas with islets of Langerhans

Abstract

Decellularization of pancreata and repopulation of these non-immunogenic matrices with islets and endothelial cells could provide transplantable, endocrine Neo- Pancreata. In this study, rat pancreata were perfusion decellularized and repopulated with intact islets, comparing three perfusion routes (Artery, Portal Vein, Pancreatic Duct). Decellularization effectively removed all cellular components but conserved the pancreas specific extracellular matrix. Digital subtraction angiography of the matrices showed a conserved integrity of the decellularized vascular system but a contrast emersion into the parenchyma via the decellularized pancreatic duct. Islets infused via the pancreatic duct leaked from the ductular system into the peri-ductular decellularized space despite their magnitude. TUNEL staining and Glucose stimulated insulin secretion revealed that islets were viable and functional after the process. We present the first available protocol for perfusion decellularization of rat pancreata via three different perfusion routes. Furthermore, we provide first proof-of-concept for the repopulation of the decellularized rat pancreata with functional islets of Langerhans. The presented technique can serve as a bioengineering platform to generate implantable and functional endocrine Neo-Pancreata.

Strategies based on organ decellularization and recellularization

“…Depending on the cell type, the first technique involves recellularization via the vascular network or another hollow structure (e.g. bile duct, ureter, intestinal lumen or airway) . Alternatively, recellularization via direct cellular injection into the parenchymal compartment is also possible.…”

“…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 .…”

Application of a novel bioreactor for in vivo engineering of pancreas tissue

“…De et al () were the first researchers to delineate the appropriateness of two‐dimensional (2D) bioscaffolds in whole‐organ engineering. Furthermore, several research groups have evaluated the capacity of in vitro techniques for whole‐organ decellularization and regeneration of pancreatic tissue, including Napierala et al (), who claimed to be the first to provide proof‐of‐concept regeneration of functional islet cells. Hereby, we demonstrate a novel islet cell repopulation technique and the successful production of acellular bioscaffolds.…”

Bioengineering the Pancreas: Cell-on-Scaffold Technology

“…Recently a group headed by Struecker has refined the entire process providing, for the first time, a proof-of-concept for the repopulation of the decellularized rat pancreas with functional islets of Langerhans [123]. Briefly rat pancreas was decellularized via vascular perfusion (1% Triton X: 0.5% SDS and 1% Triton X-100) with a flow-rate around 10 ml/min and then repopulated with approximately 2000 islets via the pancreatic duct to test viability and functionality of the islets after the process.…”