(B-42) Developing methods to improve vascularization of nephron progenitor cell grafts beneath the kidney capsule through the use of pro-angiogenic uniformly porous templated hydrogel scaffolds

Introduction:: When human kidneys are acutely injured or damaged by underlying chronic conditions, the nephrons are unable to sufficiently repair themselves and will progressively deteriorate to the point of end stage kidney disease. The fairly recent development of stem cell-derived kidney organoids has raised the possibility of achieving regenerative medicine-based therapeutic to restore renal function. However, studies in vivo have not yet achieved clinically significant integration of implanted cells with the host kidney. Here we examine the effects of implanting nephron progenitor cells (NPCs) as well as pro-angiogenic uniform porous templated hydrogel scaffolds beneath the kidney capsule as potential methods to achieve highly vascularized grafts.


Materials and Methods:: Human induced pluripotent stem cell-derived NPCs, differentiated kidney organoids, and precision porous hydrogel scaffolds were implanted beneath the kidney capsules of 8–10-week-old NOD-SCID mice. Kidneys were excised after 3-weeks and analyzed through cryosection immunofluorescent staining to look for a variety of cell markers including those that indicate major nephron segments such as podocalyxin, lotus tetragonolobus lectin and e-cadherin. Cellular material was collected by manually scraping cells from adherent cultures in a 24-well plate and aggregated through centrifugation. Hydrogels were made by filling the negative space of a sintered bead-cake mold with monomer solution, UV crosslinking the monomer, and then dissolving out the beads.


Results, Conclusions, and Discussions:: Our data shows that implantation of NPCs beneath the kidney capsule of NOD-SCID mice supports further differentiation and leads to the development of renal structures that include podocytes, and proximal and distal tubules. The developing podocytes interact with host vasculature to form chimeric glomerular structures. Additionally, implanted precision porous scaffolds allow for ubiquitous cellular infiltration and vascularization while reducing the fibrotic host response compared to similar non-porous implants. Together, the results from these experiments have demonstrated the potential for each of these two technologies to improve kidney regeneration therapies. Next steps will be to optimize cell-seeding methodology and implant combined NPC-seeded scaffolds beneath the kidney capsule to explore potential synergistic effects.


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