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Kidney Week

Abstract: TH-PO398

Controlling Proximal Nephron Arrangement and Maturity in Stem Cell-Derived Engineered Kidney Tissue

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 600 Development, Stem Cells, and Regenerative Medicine

Authors

  • Vanslambrouck, Jessica May, The Novo Nordisk Foundation Centre for Stem Cell Medicine (reNEW), Murdoch Children's Research Institute, Parkville, Melbourne, Victoria, Australia
  • Tan, Ker sin, The Novo Nordisk Foundation Centre for Stem Cell Medicine (reNEW), Murdoch Children's Research Institute, Parkville, Melbourne, Victoria, Australia
  • Mah, Sophia Ying Yii, The Novo Nordisk Foundation Centre for Stem Cell Medicine (reNEW), Murdoch Children's Research Institute, Parkville, Melbourne, Victoria, Australia
  • Dominic, Kathleen, The Novo Nordisk Foundation Centre for Stem Cell Medicine (reNEW), Murdoch Children's Research Institute, Parkville, Melbourne, Victoria, Australia
  • Little, Melissa H., The Novo Nordisk Foundation Centre for Stem Cell Medicine (reNEW), Murdoch Children's Research Institute, Parkville, Melbourne, Victoria, Australia

Group or Team Name

  • Kidney Regeneration, Stem Cell Medicine Theme.
Background

The proximal tubule (PT) structure is critical to the metabolic, endocrine, immune, and general homeostatic functions of the kidney. However, its acute vulnerability to damage and disease has made the generation of stable and accurate PTs a key objective of kidney disease research and treatment advances. While human pluripotent stem (hPSC)-derived kidney organoids are one approach, their immature PTs and disorganised nephron spatial arrangement remain a barrier to translational applications. We have previously developed PT-enhanced kidney organoids with elongated and functionalised proximal tubules. Here we report a breakthrough in the control of proximal nephron spatial arrangement achieved by exploiting biophysical parameters. This approach supports the formation of physiologically relevant S1/S2/S3 PT segmentation and is transferable to organ-on-a-chip platforms.

Methods

Standard and reporter hPSCs were differentiated to enhanced nephron progenitors prior to organoid formation (Vanslambrouck et al. Nature Protocols, 2023). Progenitors and PT-enhanced organoids were generated using standard or extrusion-based 3D cellular bioprinting techniques in combination with novel organ-on-a-chip platforms, metabolic-supportive media, and comprehensive analyses using high-content confocal imaging, transcriptional profiling, and transporter functional assays.

Results

Here we show that stringent control of organoid biophysical parameters can be used to guide the spatial arrangement of nephrons within hPSC-derived PT-enhanced kidney tissue. Using appropriate metabolic support for maturing organoid cells populations, increased levels of transporters and the formation of distinct PT cell subtypes was observed. Finally, the generation of appropriately patterned PT was possible from cryopreserved progenitors, simplifying manufacture and scale up.

Conclusion

The advances to our protocol have founded an improved PT platform likely to provide more accurate disease modelling and drug/toxicity screening opportunities.

Funding

  • Private Foundation Support