Abstract: FR-OR098

Fluidic Shear Stress Induces Vascular and Glomerular Development in Kidney Organoids

Session Information

Category: Developmental Biology and Inherited Kidney Diseases

  • 402 Stem Cells

Authors

  • Homan, Kimberly, Harvard University, Cambridge, Massachusetts, United States
  • Morizane, Ryuji, Brigham and Women's Hospital, Brookline, Massachusetts, United States
  • Gupta, Navin R., Brigham & Women''s Hospital/Massachusetts General Hospital, Brighton, Massachusetts, United States
  • Kroll, Katharina T, Harvard University, Cambridge, Massachusetts, United States
  • Kolesky, David, Harvard University, Cambridge, Massachusetts, United States
  • Skylar-Scott, Mark A, Harvard University, Cambridge, Massachusetts, United States
  • Miyoshi, Tomoya, Brigham and Women's Hospital, Brookline, Massachusetts, United States
  • Valerius, M. Todd, Brigham and Women's Hospital, Brookline, Massachusetts, United States
  • Bonventre, Joseph V., Brigham and Women's Hospital, Brookline, Massachusetts, United States
  • Lewis, Jennifer A., Harvard School of Engineering and Applied Sciences, Cambridge, Massachusetts, United States
Background

Kidney organoids, derived from human pluripotent stem cells (hPSCs), provide a novel platform to study basic kidney development, drug toxicity, and disease modeling. The cellular heterogeneity and tubular architectures recapitulated in these systems are noteworthy, and recent studies demonstrated that vascularized glomeruli can be formed with host endothelial cells upon transplantation of organoid-derived podocytes to SCID mice. However, in current organoid systems in vitro, glomerular development is imperfect and vasculature is neither perfusable nor remains viable longitudinally, limiting both the degree of relevant applications and potential extent of translatability to human physiology in vivo.

Methods

Early nephron organoids (renal vesicle stage) derived from both human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) were embedded on extracellular matrix (ECM) in 3D perfusable chips. The degree, distribution, and maturation of vascular networks were evaluated by immunostaining, RT-qPCR, and flow cytometry for FLK1, CD146, and CD31 at regular intervals when subject to variable degrees of fluidic shear stress as well as of growth factors including VEGF, as compared to controls in static chips.

Results

By subjecting renal organoids to the right combination of underlying ECM, medium components, and fluidic shear stress, the abundance of vasculature, the incidence of capillary invasion of glomerular clefts, and the number of vascularized glomerular structures as well as peritubular vasculature are significantly enhanced. We also demonstrate that the vasculature contains open lumens which can be visualized with fluorescent beads, indicating that vasculature in the organoids is perfusable.

Conclusion

Culturing renal organoids under fluidic shear stress has the potential to unlock new opportunities for glomerular disease modeling, podocyte/vascular maturation, and development of a glomerular filtration barrier in vitro.

Funding

  • NIDDK Support