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Kidney Week

Abstract: FR-OR048

Application of Cellular Extrusion Bioprinting to Improve Kidney Organoid Patterning

Session Information

  • Development and Stem Cells
    November 08, 2019 | Location: 152, Walter E. Washington Convention Center
    Abstract Time: 05:54 PM - 06:06 PM

Category: Development, Stem Cells, and Regenerative Medicine

  • 500 Development, Stem Cells, and Regenerative Medicine

Authors

  • Vanslambrouck, Jessica May, Murdoch Childrens Research Institute, Parkville, Victoria, Australia
  • Lawlor, Kynan T., Murdoch Children''s Research Institute, Parkville, Victoria, Australia
  • Higgins, James William, Organovo, Inc. , San Diego, California, United States
  • Howden, Sara E., Murdoch Childrens Research Institute, Parkville, Victoria, Australia
  • Tan, Ker sin, Murdoch Children''s Research Institute, Parkville, Victoria, Australia
  • Wilson, Sean, Murdoch Childrens Research Institute, Parkville, Victoria, Australia
  • Er, Pei Xuan, Murdoch Children's Research Institute, Parkville, Victoria, Australia
  • Combes, Alexander N., Murdoch Children's Research Institute, Parkville, Victoria, Australia
  • Arndt, Derek, Organovo, Inc. , San Diego, California, United States
  • Shepherd, Benjamin, Organovo, Inc, San Diego, California, United States
  • Chen, Alice, Organovo, Inc., San Diego, California, United States
  • Little, Melissa H., Murdoch Childrens Research Institute, Parkville, Victoria, Australia
Background

Organoids derived from induced pluripotent stem cells (iPSCs) are now being exploited as a model system for a variety of organs, especially for structurally complex organs such as the kidney. iPSC-derived kidney organoids show great potential for modelling kidney diseases and improving our understanding of disease pathogenesis. However, they do not yet accurately recapitulate the cellular maturity and compartmental organisation of the human kidney in vivo. Changes in culture format may allow for improved patterning by altering biophysical properties, cell-cell interactions and growth factor gradients. Here we report the application of 3D cellular extrusion bioprinting to i) improve quality control and ii) modify organoid morphogenesis and maturation.

Methods

A series of fluorescent reporter iPSC lines designed to report the formation of specific cellular compartments of the kidney, such as podocytes, proximal tubule and distal nephron/ureteric epithelium, were subjected to monolayer differentiation as previously described (Takasato et al, 2016). Generation of organoids was performed using extrusion-based NovoGen Bioprinter® MMX technology (Organovo), with variations in cell density and printing configuration, and compared to manually pelleted organoids. Organoids were analysed via live imaging for fluorescent reporters, confocal immunofluorescence and quantitative image analysis.

Results

Bioprinting facilitated rapid, uniform and highly reproducible organoid production. Organoids were well-patterned and comprised all kidney cell types and compartments previously described using the manual technique. Changes in cell density, organoid shape and thickness also modified nephron number, patterning and the formation of off-target populations.

Conclusion

Here we demonstrate that biophysical properties of organoids at the time of their generation can have a significant impact of their morphogenesis and differentiation, potentially affecting cellular maturity and differentiation trajectory. Further research into the variables of organoid generation will be fundamental to achieve and maximise future downstream applications.

Funding

  • NIDDK Support