Abstract: FR-PO977

Development of Functional Vasculature in Decellularized Whole Porcine Kidneys with Human Endothelial Cells

Session Information

Category: Bioengineering and Informatics

  • 101 Bioengineering and Informatics

Authors

  • Cabral, Pablo D., Miromatrix Medical Inc., Eden Prairie, Minnesota, United States
  • Seetapun, Dominique, Miromatrix Medical Inc., Eden Prairie, Minnesota, United States
  • Atputhanathan, Senthuran, Miromatrix Medical Inc., Eden Prairie, Minnesota, United States
  • Ross, Jeff, Miromatrix Medical, Eden Prairie, Minnesota, United States
  • Black, Morgan D., Miromatrix Medical, Inc., Eden Prairie, Minnesota, United States
Background

End stage renal disease (ESRD) represents a major epidemic both in the US and worldwide. Patients with ESRD have two options: kidney transplantation or hemodialysis. Due to the organ donor shortage, only a few patients receive transplants. The alternative, hemodialysis, has a 5-year mortality rate of 35% compared to only 3% for transplantation. With ~500,000 hemodialysis patients in the US, a new therapy is needed. The ultimate solution would be a bioengineered kidney to solve the chronic shortage. A critical first step is the demonstration of a functional vasculature to continuously perfuse blood. We demonstrate that a functional vasculature can be achieved by repopulating decellularized kidneys with human primary cells.

Methods

Whole porcine kidneys were perfusion decellularized without compromising the native microarchitecture of both the vascular and tubular compartments. Human umbilical vein endothelial cells (HUVEC) and primary human epithelial cells (HRE) were perfused into the vascular and tubular compartments respectively. Whole organ culture was performed under continous perfusion and key metabolic parameters were monitored daily to assess cell proliferation and viability.
Functional vasculature assessment was performed with blood loops using whole porcine blood to model in-vivo performance.

Results

Cellular engraftment and viability were measured by metabolic parameters including glucose consumption over 3 to 4 weeks (n=12) to achieve the desired level. These data were further corroborated by histological analysis of formalin fixed kidney sections demonstrating the presence of a single layer of engrafted cells on vascular and tubular compartments. Furthermore, cells positive for the endothelial cell marker CD31 were confined to the vascular compartment and cells positive for the epithelial cell marker e-cadherin were confined to the tubular compartment. Vascular functionality was characterized by blood loops and demonstrated long-term continuous perfusion of whole blood compared at physiological pressures compared to non-recellularized kidneys that demonstrated the lack of flow after a few minutes.

Conclusion

These results demonstrate the ability to generate a functional vasculature in recellularized kidney grafts, a critical first step in the engineering of a fully bioengineered kidney.

Funding

  • Commercial Support