Recellularization of whole porcine kidneys with human epithelial and endothelial cells
Nafiseh Poornejad, Evan M. Buckmiller, Jaron J. Lundwall, Ho Hin Ma, Beverly L. Roeder, Alonzo D. Cook
Brigham Young University, Provo, Utah
Whole organ engineering is a promising alternative for organ transplantation. Murine, porcine and human kidneys have been successfully decellularized and rat kidneys have previously been recellularized with human umbilical vein endothelial cells (HUVECs), rat neonatal kidney cells, and murine stem cells [1, 2]. In this study previous technology was scaled up to porcine kidneys as a model of human-sized kidneys. Decellularization of porcine kidneys was optimized using pressure control and tonic cycles and the effects of freezing/thawing conditions on decellularized extracellular matrix were investigated . Sterilization methods for whole kidneys were also optimized to improve cell adhesion and proliferation. Human renal cortical tubular epithelium (RCTEs) cells and murine endothelial cells (MS-1) were used to repopulate the epithelial and endothelial niches of renal tissue, respectively. More than 6×108 endothelial or epithelial cells were perfused through the whole decellularized porcine kidneys for 30 min, either through the ureter for epithelial cells or through the artery for endothelial cells, and then media perfusion was continued for at least 7 days. Immunohistological staining demonstrated that tubules were repopulated with epithelial cells and endothelial cells were located in the complicated vasculature of the kidney. Metabolic activity of reseeded RCTEs was determined by resazurin reduction assay and glucose assay kit. The epithelial cells activity throughout the kidney will be checked by creatinine clearance measurement. This study demonstrated the feasibility of regeneration of whole human-sized kidneys using decellularized porcine kidneys and human cells.
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