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2015 Abstracts

Determining the Integrity of Decellularized Porcine Kidney Scaffolds

Benjamin Buttars, Jeffrey Nielson, Spencer Baker, Jonathon Thibaudeau, Angela Nakalembe, Tim
Frost, Blake Cannon, Robert Fuller, Brinden Elton, Daniel Scott, Nafiseh Poornejad,
and Cameron Bruner, Brigham Young University

Engineering

Chronic kidney disease can be treated by organ transplantation, but the number of patients on the waiting list far exceeds the number of available donors. The patients who are fortunate enough to receive an organ need to stay on immunosuppressive medication for the remainder of their lives. Non-immunogenic tissue engineered organs could replace donor organ transplantation. We hypothesize that by seeding autologous cells on an appropriate acellular scaffold we may be able to create an inexhaustible supply of non-immunogenic organs and avoid organ rejection. In this study we have used porcine kidneys as a comparative model of human kidneys. Decellularization was used to create an intact, acellular, non-immunogenic collagenous scaffold structure. We used chelating agents and osmotic shock (alternating hypotonic, hypertonic and sodium dodecyl sulfate solutions) combined with SDS to reduce detergent exposure time by 6-fold compared to published methods using only ionic and non-ionic detergents. DNA assay results demonstrated essentially complete cell removal in all applied methods (Total DNA < 50 ng/mg). Collagen and glycosaminoglycans, which are essential for cell proliferation, differentiation, and adhesion to the scaffold, were visualized to compare the efficacy of structure integrity after decellularization. Our results showed increased integrity of the scaffold with decreasing detergent exposure time. Also, the cell adhesion and proliferation capacities of the acellular scaffolds were examined using mouse endothelial cells expressing green fluorescent protein (GFP-MS1). The resulting scaffolds demonstrated an increased capacity for cell attachment, growth and differentiation. This research demonstrates the potential for creating non-immunogenic tissue-engineered organs.