Decellularization and Recellularization of Rabbit Kidney Using Adipose-Derived Mesenchymal Stem Cells for Renal Tissue Engineering

Abstract

The effectiveness of decellularization and recellularization of rabbit kidney scaffolds with autologous adipose-derived mesenchymal stem cells (ADMSCs) was assessed. The static-based technique was judged against the perfusion-based method both in decellularization and recellularization processes. Kidneys were obtained from 8 rabbits and divided into two groups. Two kidneys did not undergo any processing and were served as the control group. Kidneys of group I (N = 7) were cannulated via the renal artery and decellularized with a detergent-based method. We reseeded these constructs with ADMSCs using a peristaltic pump. The static method was applied to decellularize and recellularize thin slices of the seven kidneys (group II, N = 7) using the L929 cell line. Several tests were performed to evaluate the efficacy of the decellularization and recellularization processes. All cellular and nuclear constituents were eliminated from the scaffolds of both groups confirmed with hematoxylin and eosin (H&E), trichrome, 4′,6-diamidino-2-phenylindole (DAPI) staining, and DNA quantification. Cell viability was over 89% as performed by MTT assay. Cells were effectively seeded in scaffolds of both groups, with a superior arrangement and a higher concentration in the perfusion-based technique. Immunohistochemistry (IHC) staining demonstrated preservation of native expression patterns of extracellular matrix proteins with better results in perfusion-based technique. The results suggest that organs with the distributed micro-vasculature system (perfusion method) may benefit from being engineered with the dynamic procedure compared with organs with empty chambers and poor vasculature system (static system). Previously, we evaluated the efficacy of the transplantation of decellularized kidneys in a rat model. In this study, we try to assess the efficacy of two decellularization and recellularization protocols of kidney organs for further renal tissue engineering. We also aim to produce renal scaffolds with structural, mechanical, and physiological characteristics that are crucial for engineering main renal structures, using adipose-derived mesenchymal stem cells. Finding the most suitable and feasible decellularization and recellularization methods is important in several medicinal therapeutics. In the future, we will emphasize on the long-term viability and functionality of obtained kidney scaffolds in clinically translatable applications. We will also compare different cell lineages to obtain the most efficient scaffold for further renal tissue engineering.