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Abstract: TH-PO0532

Perfusable Three-Dimensional (3D) Models of Ureteric Bud and Collecting Duct Tubules

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 600 Development, Stem Cells, and Regenerative Medicine

Authors

  • van Gaal, Ronald C., Harvard University, Cambridge, Massachusetts, United States
  • Wolf, Kayla J., Harvard University, Cambridge, Massachusetts, United States
  • Rubins, Jonathan Eric, Harvard University, Cambridge, Massachusetts, United States
  • Kroll, Katharina T., Harvard University, Cambridge, Massachusetts, United States
  • Satlin, Lisa M., Icahn School of Medicine at Mount Sinai, New York, New York, United States
  • Lewis, Jennifer A., Harvard University, Cambridge, Massachusetts, United States
Background

Engineered kidney tissues are limited in part by a lack of collecting duct network and ureter to facilitate urine drainage and modification. Although protocols to derive ureteric bud (UB) and subsequently collecting duct organoids directly from human pluripotent stem cells (hPSCs) have recently been developed, these organoids are not perfusable, lacking both biophysical cues from luminal flow and a drainage outlet. Here, we develop a perfusable model of the UB and collecting duct derived from hPSCs.

Methods

To fabricate our model, dissociated UB organoids were seeded into a 3D perfusable channel embedded within optimized extracellular matrix containing basement membrane matrix and collagen-I. We then differentiate UB tubules on-chip to collecting ducts in the presence of luminal flow. As a first step toward rapidly generating extensive tubular networks that can connect with a drainage outlet in bioengineered tissue, we bioprint UB cells adjacent to the perfusable UB channel and demonstrate fusion of the printed network to the drainage channel.

Results

These UB cells on-chip form a confluent monolayer and maintain UB-like marker expression and morphology over several days to weeks. Notably, UB cells in the monolayer bud into the surrounding matrix while maintaining luminal interconnection with the central perfusable lumen resembling early branching morphogenesis observed in developing kidneys. We find that on chip collecting duct differentiation resulted in decreased UB markers and increased collecting duct markers relative to UB tubules. Moreover, we found differential marker expression is enhanced by luminal flow relative to static controls.

Conclusion

This platform can facilitate fundamental understanding of development and disease in human collecting ducts and ultimately serve as a drainage channel upon integration with biomanufactured tissue.

Funding

  • Other NIH Support

Digital Object Identifier (DOI)