The aim of this study was to acquire an effective method for preparation of rat decellularized kidney scaffolds capable of supporting proliferation and differentiation of human adipose tissue derived mesenchymal stem cells (AD-MSCs) into kidney cells. We compared two detergents, the sodium dodecyl sulfate (SDS) and triton X-100 for decellularization. The efficiency of these methods was assessed by Hematoxylin and Eosin (H&E), 4\\\\\\\', 6 diamidino-2-phenylindole and immunohistochemistry (IHC) staining. In the next step, AD-MSCs were seeded into the SDS-treated scaffolds and assessed after three weeks of culture. Proliferation and differentiation of AD-MSCs into kidney-specific cell types were then analyzed by H&E and IHC staining. The histological examinations revealed that SDS was more efficient in removing kidney cells at all-time points compared to triton X-100. Also, in the SDS-treated sections the native extracellular matrix was more preserved than the triton-treated samples. Laminin was completely preserved during decellularization procedure using SDS. Cell attachment in the renal scaffold was observed after recellularization. Furthermore, differentiation of AD-MSCs into epithelial and endothelial cells was confirmed by expression of Na-K ATPase and vascular endothelial growth factor receptor 2 (VEGFR-2) in seeded rat renal scaffolds, respectively. Our findings illustrated that SDS was more effective for decellularization of rat kidney compared to triton X-100. We presented an optimized method for decellularization and recellularization of rat kidneys to create functional renal natural scaffolds. These natural scaffolds supported the growth of AD-MSCs and could also induce differentiation of these cells into epithelial and endothelial cells.

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