Decellularization and Recellularization of Rat Livers With Hepatocytes and Endothelial Progenitor Cells

Abstract

Whole-organ decellularization has been identified as a promising choice for tissue engineering. The aim of the present study was to engineer intact whole rat liver scaffolds and repopulate them with hepatocytes and endothelial progenitor cells (EPCs) in a bioreactor. Decellularized liver scaffolds were obtained by perfusing Triton X-100 with ammonium hydroxide. The architecture and composition of the original extracellular matrix were preserved, as confirmed by morphologic, histological, and immunolabeling methods. To determine biocompatibility, the scaffold was embedded in the subcutaneous adipose layer of the back of a heterologous animal to observe the infiltration of inflammatory cells. Hepatocytes were reseeded using a parenchymal injection method and cultured by continuous perfusion. EPCs were reseeded using a portal vein infusion method. Morphologic and functional examination showed that the hepatocytes and EPCs grew well in the scaffold. The present study describes an effective method of decellularization and recellularization of rat livers, providing the foundation for liver engineering and the development of bioartificial livers.

Culture of iPSCs Derived Pancreatic β-Like Cells In Vitro Using Decellularized Pancreatic Scaffolds: A Preliminary Trial

“…Secondly, our research demonstrated that transplanted cells did not distribute uniformly, although vascular perfusion and multipositional parenchymal injection were taken simultaneously. Also revascularization remains a major bottleneck towards biological engineering, and while we have promising preliminary data towards addressing this issue, more works still need to be performed in future studies [45]. …”

Recellularization via the bile duct supports functional allogenic and xenogenic cell growth on a decellularized rat liver scaffold

“…Two major obstacles to creating a transplantable bioengineered liver are complete re-endothelialization of the vascular tree 17 and growing both parenchymal and non-parenchymal cells on the same ECM scaffolding with appropriate organization and cell compartment localization. 10 Endothelialization of the vasculature is necessary to prevent thrombosis during transplantation as any exposed collagen will activate platelets and the extrinsic coagulation cascade making successful transplantation impossible.…”

“…2 Following the success of perfusion decellularization in the cardiac model, the principle has been applied to other whole organs such as the kidney, [3][4][5][6] lungs, [7][8][9] and liver. 6,[10][11][12][13][14][15][16][17][18] A number of effective strategies for the decellularization of rat livers have been described by Uygun et al, 10 Bao et al 19 and Baptista et al…”

“…20 However, the ability to reproducibly generate a robust recellularized construct has proven elusive. 7,11,17,21 Early studies attempted to seed the scaffold with primary hepatocytes, and these efforts were followed by the use of stem and progenitor cells. 10,11,17,21,22 More recent work has demonstrated the importance of cell-to-cell interactions, thus highlighting the importance of multiple cell lineages in recreating a functional native liver.…”

“…7,11,17,21 Early studies attempted to seed the scaffold with primary hepatocytes, and these efforts were followed by the use of stem and progenitor cells. 10,11,17,21,22 More recent work has demonstrated the importance of cell-to-cell interactions, thus highlighting the importance of multiple cell lineages in recreating a functional native liver. 21 Transplantation of the recellularized construct has resulted in rapid graft thrombosis, likely secondary to exposed collagen in the deendothelialized vasculature.…”