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Bioprinted pluripotent stem cell-derived kidney organoids provide opportunities for high content screening

J. William Higgins, Alison Chambon, Kristina Bishard, Anke Hartung, Derek Arndt, Jamie Brugnano, Pei Xuan Er, Kynan T. Lawlor, Jessica M. Vanslambrouck, Sean Wilson, Alexander N. Combes, Sara E. Howden, Ker Sin Tan, Santhosh V. Kumar, Lorna J. Hale, Benjamin Shepherd, Stephen Pentoney, Sharon C. Presnell, Alice E. Chen, Melissa H. Little
doi: https://doi.org/10.1101/505396
J. William Higgins
1Organovo Inc, San Diego
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Alison Chambon
1Organovo Inc, San Diego
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Kristina Bishard
1Organovo Inc, San Diego
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Anke Hartung
1Organovo Inc, San Diego
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Derek Arndt
1Organovo Inc, San Diego
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Jamie Brugnano
1Organovo Inc, San Diego
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Pei Xuan Er
2Murdoch Children’s Research Institute, Flemington Rd, Parkville, VIC, Australia
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Kynan T. Lawlor
2Murdoch Children’s Research Institute, Flemington Rd, Parkville, VIC, Australia
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Jessica M. Vanslambrouck
2Murdoch Children’s Research Institute, Flemington Rd, Parkville, VIC, Australia
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Sean Wilson
2Murdoch Children’s Research Institute, Flemington Rd, Parkville, VIC, Australia
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Alexander N. Combes
2Murdoch Children’s Research Institute, Flemington Rd, Parkville, VIC, Australia
3Department of Anatomy and Neuroscience, The University of Melbourne, VIC, Australia.
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Sara E. Howden
2Murdoch Children’s Research Institute, Flemington Rd, Parkville, VIC, Australia
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Ker Sin Tan
2Murdoch Children’s Research Institute, Flemington Rd, Parkville, VIC, Australia
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Santhosh V. Kumar
2Murdoch Children’s Research Institute, Flemington Rd, Parkville, VIC, Australia
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Lorna J. Hale
2Murdoch Children’s Research Institute, Flemington Rd, Parkville, VIC, Australia
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Benjamin Shepherd
1Organovo Inc, San Diego
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Stephen Pentoney
1Organovo Inc, San Diego
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Sharon C. Presnell
1Organovo Inc, San Diego
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Alice E. Chen
1Organovo Inc, San Diego
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Melissa H. Little
2Murdoch Children’s Research Institute, Flemington Rd, Parkville, VIC, Australia
3Department of Anatomy and Neuroscience, The University of Melbourne, VIC, Australia.
4Department of Paediatrics, The University of Melbourne, VIC, Australia.
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  • For correspondence: melissa.little@mcri.edu.au
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Abstract

Recent advances in the directed differentiation of human pluripotent stem cells to kidney brings with it the prospect of drug screening and disease modelling using patient-derived stem cell lines. Development of such an approach for high content screening will require substantial quality control and improvements in throughput. Here we demonstrate the use of the NovoGen MMX 3D bioprinter for the generation of highly reproducible kidney organoids from as few as 4,000 cells. Histological and immunohistochemical analyses confirmed the presence of renal epithelium, glomeruli, stroma and endothelium, while single cell RNAseq revealed equivalence to the cell clusters present within previously described organoids. The process is highly reproducible, rapid and transferable between cell lines, including genetically engineered reporter lines. We also demonstrate the capacity to bioprint organoids in a 96-well format and screen for response to doxorubicin toxicity as a proof of concept for high content compound screening.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted December 23, 2018.
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Bioprinted pluripotent stem cell-derived kidney organoids provide opportunities for high content screening
J. William Higgins, Alison Chambon, Kristina Bishard, Anke Hartung, Derek Arndt, Jamie Brugnano, Pei Xuan Er, Kynan T. Lawlor, Jessica M. Vanslambrouck, Sean Wilson, Alexander N. Combes, Sara E. Howden, Ker Sin Tan, Santhosh V. Kumar, Lorna J. Hale, Benjamin Shepherd, Stephen Pentoney, Sharon C. Presnell, Alice E. Chen, Melissa H. Little
bioRxiv 505396; doi: https://doi.org/10.1101/505396
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Bioprinted pluripotent stem cell-derived kidney organoids provide opportunities for high content screening
J. William Higgins, Alison Chambon, Kristina Bishard, Anke Hartung, Derek Arndt, Jamie Brugnano, Pei Xuan Er, Kynan T. Lawlor, Jessica M. Vanslambrouck, Sean Wilson, Alexander N. Combes, Sara E. Howden, Ker Sin Tan, Santhosh V. Kumar, Lorna J. Hale, Benjamin Shepherd, Stephen Pentoney, Sharon C. Presnell, Alice E. Chen, Melissa H. Little
bioRxiv 505396; doi: https://doi.org/10.1101/505396

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