Abstract: PO0310
Ex Vivo Perfusion and Initial Function of a Recellularized Human-Scale Bioengineered Kidney
Session Information
- Bioengineering
October 22, 2020 | Location: On-Demand
Abstract Time: 10:00 AM - 12:00 PM
Category: Bioengineering
- 300 Bioengineering
Authors
- Uzarski, Joseph S., Miromatrix Medical, Inc., Eden Prairie, Minnesota, United States
- Beck, Emily C., Miromatrix Medical, Inc., Eden Prairie, Minnesota, United States
- Russell, Emily, Miromatrix Medical, Inc., Eden Prairie, Minnesota, United States
- Jaffe, James, Miromatrix Medical, Inc., Eden Prairie, Minnesota, United States
- Shapiro, Ron, Recanati/Miller Transplantation Institute at Mount Sinai, New York, New York, United States
- Florman, Sander, Mount Sinai Health System, New York, New York, United States
- Davidow, Dominique, Miromatrix Medical, Inc., Eden Prairie, Minnesota, United States
- Gilbert, Thomas, Miromatrix Medical, Inc., Eden Prairie, Minnesota, United States
- Ross, Jeff, Miromatrix Medical, Inc., Eden Prairie, Minnesota, United States
Background
The need for more transplantable kidneys is greater than ever, with nearly 100,000 patients actively waiting for a kidney. To meet this growing demand, our team is developing a fully transplantable bioengineered kidney (BEK) by seeding cells into perfusion-decellularized porcine kidney scaffolds. Previously, orthotopic transplantation of HUVEC-only BEKs in pigs resulted in 83.3% (n=5/6 pigs) renal perfusion at 7 days post in vivo implantation. Building on these results, the focus of the current study was to recellularize the glomerulus of a clinically relevant whole kidney matrix and then assess the preliminary filtration function.
Methods
Adult porcine kidneys were decellularized via detergent perfusion through the vasculature. Primary glomerular cells were isolated from fresh porcine kidneys or rejected human kidneys. The porcine matrix was then seeded with either human umbilical vein endothelial cells (HUVECs), HUVECs and porcine glomerular cells as a model system, or HUVECS and human glomerular cells. The recellularized grafts were then cultured using a custom perfusion recellularization bioreactor until sufficient cellular coverage of the vasculature was obtained through nondestructive metabolic markers. Both HUVEC-only and co-culture BEKs were then implanted in an ex vivo blood loop for 30 minutes, where ureter effluent was collected and analyzed for filtration function.
Results
Sufficient histological vascular coverage with endothelial cells and thromboresistance with vascular patency was found for grafts with a minimum glucose consumption rate of 20 mg/hr. From the ex vivo blood loop test, the ureter effluent hematocrit concentration in the co-culture BEK was found to be undetectable. In comparison, the endothelial-only BEK and the pig’s blood hematocrit levels were both 22%. Finally, the addition of glomerular cells in the BEK restored physiological flow rates of ureter effluent.
Conclusion
These results demonstrated human cellular engraftment and growth, long-term vascular patency, sustained hemoperfusion, removal of processed filtrate, and early signs of filtration and waste clearance in BEKs, which moves the field closer to an alternative for kidney transplantation.
Funding
- Commercial Support –