Recellularization of Well-Preserved Rat Acellular Kidney Scaffold Using Mesenchymal Stem Cells

Background and Aim: The development of tissue engineering havecatalyzed due to a gap between limited organ supply and increasingrequests for patients with the end-stage renal disease. The choice anappropriate scaffold for cell seeding and production of the functionalorgan for transplant purpose is the critical challenges in tissue engineering.An important technique in regenerative medicine to prepare an acellularECM is the decellularization of native tissues. The purpose of this studywas to acquire effective method for preparation of decellularized kidneyscaffold which can proliferate and differentiate of MSCs into kidney cells.Methods: After removing the kidney, the adipose tissue and the capsulearound the kidney were removed. Kidney sections were washed twicewith phosphate buffered saline (PBS), followed by decellularization in asolution of either 1% Triton X-100 or sodium dodecyl sulfate 1% (SDS).The decellularization solution was changed 4 hours after initial tissueharvest and then every 24 hours until tissues were transparent (for 14days). In order to confirmation of decellularization, hematoxylin-eosin(H&E) and DAPI staining were performed on days 2, 5, 10, and 14. Then,we seeded 2×105 hAd-MSCs onto time sufficient and suitable sterilekidney scaffold to allow attachment of cells, 1 hour after seeding 1 mLDMEM was added to any of wells, and they incubated at 37℃ and 5%CO2 for 3 weeks. H&E staining was performed for measure cell viabilityand proliferation of cells within the renal scaffold.Results: DAPI staining approved the SDS-treated sections were moredecellularized than the Triton-treated sections at all times. The results ofhematoxylin and eosin staining revealed that in the SDS-treated sectionsthe native ECM architecture, integration of renal vascular and glomerularstructures was more preserved than the Triton-treated sections. Moreover,migration and establishment of a number of cells to the renal scaffoldwere observed after recellularization. In addition, cell accumulation onthe scaffold surface as well as the migration of the cells to the depth ofkidney-formed an epithelium-like structure.Conclusion: Our methods can successfully decellularize rat kidneys toproduce functional renal ECM scaffolds. These scaffolds maintain theirbasic components, retain intact vasculature and show promise for kidneyregeneration.

Keywords:

Kidney , Extracellular matrix scaffold , Decellularization , Recellularization , Mesenchymal stem cells