Decellularized Kidney Scaffold Mediated Renal Regeneration in a Rhesus Macaque Mulatta Monkey Model

Abstract

Introduction: For regeneration of complex organs, such as the kidney, decellularized (DC) tissues are promising candidates that provide a neutral environment.<br /> <br /> Objective: We successfully DC rhesus monkey kidneys by perfusion of trypsin/EDTA, sodium dodecyl sulfate (SDS), triton X-100, and peracetic acid/ethanol. Subsequently, we analysed the samples using quantitative and qualitative analysis.<br /> <br /> Methods: Histological staining, DNA quantification and scanning electron microscopy (SEM) was used to investigate cellular removal evaluation. Also, ECM architecture integrity of vascular tree was assessed by x-ray fluoroscopy. The biocompatibility properties of the DC kidney tissues were evaluated by the culture of human embryonic kidney 293 cells (HEK). To evaluate the renal function recovery, 30% of DC tissue was partially grafted into 70% of nephrectomized monkey kidney.<br /> <br /> Results: Perfusion-decellularization monkey kidneys retained their essential ECM architecture, intact vascular tree, and cellular compatibility ensured clearance of cellular material, which directly impacts immunoreactivity during transplantation. Quantitative assay and immunohistochemistry demonstrated preservation of native expression patterns and integrity of ECM components, including glycosaminoglycan (GAG), collagen, fibronectin, elastin, and laminin. Also, x-ray fluoroscopy confirmed the integrity and patency of vascular network. Sonography and CT scan images showed significant recovery of renal function after partial transplantation. Histological analysis confirmed increases in renal size and regeneration.<br /> <br /> Conclusion: Monkey kidneys could be efficiently decellularized by our novel perfusion-decellularization protocol that can be scaled up for use in human organs. This method represents a step towards development of a transplantable organ using tissue engineering techniques.