Date: Saturday, May 30, 2020
Session Time: 3:15pm-4:00pm
Presentation Time: 3:30pm-4:00pm
*Purpose: Our vision is to build a human kidney by combining human stem cells with porcine kidney extracellular matrix.
*Methods: Kidneys were decellularized followed by repopulation with cells and then placed in bioreactors that were designed in our lab. Induced pluripotent stem cells derived from patients were made using the Epi5 reprogramming system. Cell factories were employed to grow large numbers of cells. The function of the manufactured kidneys were measured using the Normothermic Ex Vivo Kidney Perfusion (NEVKP) System.
*Results: Our goal here was to develop a seamless work-flow for kidney production from stem cells. We optimized the kidney decellularization process, optimized iPSC production and differentiation to renal cells, tested methodology for large-scale cell manufacturing, built and tested bioreactors capable of supporting recellularized kidney growth and differentiation, and demonstrate that the NEVKP system can be used to measure the function of kidneys. Mass Spectrometry, Protein arrays and GAG assays were used to determine the optimized decellularization protocol for mouse and porcine kidneys. Detergent concentrations at or below the critical micelle concentration were successful to remove native cells while reducing the loss of key extracellular matrix (ECM) proteins. Specialized bioreactors were designed and manufactured to maintain organ survival of native or recellularized organs. The mouse and porcine kidney bioreactors were successfully tested with native kidney growth and for loading cells into decellularized kidneys using negative pressure through the ureter. Cell/organ survival after 10 days was achieved. iPSC were produced from human urine samples (n=6), Umbilical Cord Blood (n=2) and Umbilical Cord Tissue MSC (n=10). Testing for function of decellularized and recellularized porcine kidneys using the NEVKP system revealed that the ECM remained intact and the kidney could be perfused without damage. Minimal function was recorded over a short time (t=15 min).
*Conclusions: There are multiple steps that are essential in building a kidney. To be successful a team of scientists, engineers and transplant surgeons are required. In this study we successfully demonstrated that a seamless workflow encompassing multiple disciplines in multiple labs could be executed, thus demonstrating the feasibility of producing patient matched organs for transplantation.
To cite this abstract in AMA style:Rogers IM, Whiteley J, Chow T, Bongolan T, Mazilescu L, Urbanellis P, Ganesh S, Selzner M. Building Kidneys from Stem Cells for Transplantation [abstract]. Am J Transplant. 2020; 20 (suppl 3). https://atcmeetingabstracts.com/abstract/building-kidneys-from-stem-cells-for-transplantation/. Accessed June 15, 2021.
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