Tissue Engineering of Well Preserved Acellular Rat Kidney Scaffold Using of Mesenchymal Stem Cells

Document Type : Original Article

Authors

1 Department of Physiology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

3 Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

4 Urology and Nephrology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

5 2. Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

6 Department of Physiology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

10.30466/vrf.2019.104640.2491

Abstract

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 Adipose tissue derived mesenchymal stem cell (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 (DAPI) and immunohistochemistry (IHC) stainings. In the next step, Ad-MSCs were seeded into the SDS-treated scaffolds were assessed after 3 weeks of culture. Proliferation and differentiation of Ad-MSCs into kidney-specific cell types on these scaffolds were then analyzed by H&E and IHC stainings. The histological examinations revealed that SDS was more efficient in removing kidney cells at all times plants as compared with triton X-100, in the SDS-treated sections the native extracellular matrix was more preserved than the triton-treated sections. Laminin was completely preserved during decellularization procedure using SDS. Moreover, cell attachment in the renal scaffold was observed after reccelularization. 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 detergent for decellularization of rat kidney compared with triton X-100 and show 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 induce differentiation into epithelial and endothelial cells.

Keywords



Articles in Press, Accepted Manuscript
Available Online from 14 June 2021
  • Receive Date: 03 March 2019
  • Revise Date: 27 August 2019
  • Accept Date: 14 September 2019