Abstract: TH-PO414
Induced Kidney Progenitor Cells Provide a Rapid One-Pot Starting Material for Nephron Regeneration
Session Information
- Development, Organoids, Injury, and Regeneration
October 24, 2024 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 600 Development, Stem Cells, and Regenerative Medicine
Authors
- Vincent, Thomas, University of Washington, Seattle, Washington, United States
- Namestnikov, Michael, Tel Aviv University, Tel Aviv, Tel Aviv, Israel
- Dekel, Benjamin, Sheba Medical Center, Tel Hashomer, Tel Aviv, Israel
- Freedman, Benjamin S., University of Washington, Seattle, Washington, United States
Background
For human kidney embryonic rudiment transplantation, there is an early window of opportunity that promotes enhanced graft formation. Protocols to differentiate kidney organoids from human stem cells mimics in vivo organogenesis as cells progress through intermediate mesoderm, metanephric mesenchyme, and progenitor cell stages. This raises the possibility of achieving tissue engineered therapeutics to restore renal function. We therefore tested the impact of early organoids on patterns of in vivo engraftment.
Methods
Organoids were differentiated from stem cells using a brief CHIR pulse. Cells were characterized temporally through qPCR and immunofluorescence staining. Implanted material was collected at multiple timepoints (days 4, 7, and 15) by scraping cells from an adherent 24-well plate. These pellets were inserted beneath the kidney capsules of NOD-SCID mice. Kidneys were excised after 3-weeks and analyzed through immunofluorescent staining.
Results
Here we show that 4 days of differentiation generates an induced metanephric mesenchyme-like population. When implanted beneath the kidney capsule, they develop into well-vascularized glomeruli containing human podocytes and GATA3+, PDGFRβ+ mesangial cells contiguous with tubular structures (Fig 1). Implanting of d7 cells consistently produced grafts with more PODXL+ and LTL+ structures than d15 late-stage kidney organoid implantations (Fig 2).
Conclusion
These results suggest using progenitor cells as starting material may have advantages in the development of future therapeutics over late-stage organoids. We are currently working to use these engraftments to study drug delivery and create novel disease models. In the future, we hope to address challenges related to distal tubule connections in order to achieve successful urine drainage from the graft and begin to assess function.
Funding
- NIDDK Support