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Abstract: TH-OR131

Kidney Organoids Generated through Heterochronic Recombination of Progenitors Show Nephron Segmentation and Vascular Connection with Host In Vivo

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 501 Development, Stem Cells, and Regenerative Medicine: Basic

Authors

  • Gupta, Ashwani Kumar, Maine Medical Center Research Institute, Scarborough, Maine, United States
  • Karolak, Michele Jean, Maine Medical Center Research Institute, Scarborough, Maine, United States
  • Oxburgh, Leif, Maine Medical Center Research Institute, Scarborough, Maine, United States
Background

In recent years procedures have been developed to differentiate human induced pluripotent stem cells (hiPSC) to renal tissues. In the present study, we investigated the potential of heterochronic recombination of hiPSC-derived kidney progenitors to increase tubular density in hiPSC-derived kidney organoids. Characterization of this technique included organoid engraftment, nephron segmentation, graft vascularization and perfusion in vivo.

Methods

hiPSC-derived kidney progenitors were treated with a WNT/β-catenin agonist for two days to induce epithelialization and were mixed with fresh progenitors, aggregated and cultured at the air-liquid interface. On day 9, these organoids were engrafted under the kidney capsule of NSG mice. Grafts were harvested after 3 weeks and nephron segmentation, vascularization and perfusion were evaluated.

Results

Heterochronic recombination of kidney progenitors induced robust tubulogenesis in kidney organoids compared with unmixed cells. Tubules stained with molecular markers for proximal and distal tubules and revealed segmentation. Podocytes (Podxl+), stromal cells (Meis1+) and endothelial cells (CD31+) were abundant. Engrafted organoids showed differentiation of complex graft-derived glomeruli with vascular networks (CD31+ Endomucin+), podocytes (Podxl+ WT1+), mesangial cells (Pdgfrβ+), glomerular basement membrane (Laminin+ Col I+ Col IV+), and juxtaglomerular cells (Renin+). Proximal tubules (E cadherin- LTL+), thick ascending limb of the loop of Henle (Tamm-Horsfall protein+), distal tubule (BRN1+), connecting tubule (E cadherin+ GATA3- DBA+), collecting duct (Troma-1+ GATA3+ DBA+), and stromal cells (Meis1+ Pdgfrβ+) were also present in the grafts. Engrafted organoids showed vascularization (CD31+ Endomucin+ SMA+) and direct connections with host vasculature. All nephron structures and stromal cells in the graft were iPSC derived (HuNu+), whereas endothelial cells were derived from the host (HuNu-).

Conclusion

Heterochronic recombination of kidney progenitors results in vigorous differentiation of human iPSC-derived kidney tissue in vitro and in vivo.

Funding

  • NIDDK Support