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Kidney Week

Abstract: FR-PO768

Optimizing Human Kidney Organoids for Modeling Nephrotoxicity, Kidney Injury, and Kidney Diseases and for Screening to Identify Therapeutic Options

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 500 Development, Stem Cells, and Regenerative Medicine

Authors

  • Lee, Nathan, Brigham and Women''s Hospital, Boston, Massachusetts, United States
  • Choi, Kyung hwa, Brigham and Women''s Hospital, Boston, United States
  • Zeleznik ramuta, Taja, Brigham and Women''s Hospital, Boston, United States
  • Bonventre, Joseph V., Brigham and Women's Hospital, Boston, Massachusetts, United States
Background

To optimize their potential in modeling nephrotoxicity and kidney disease, pluripotent stem cell-derived kidney organoid protocols are being optimized for enhancing reproducibility, differentiation, scalability and reducing costs.

Methods

Organoids were generated by modifications of our laboratory’s prior published techniques. Importantly, organoids were generated without use of undefined matrices. Cell types and structures were identified by immunostaining for LTL, PODXL, NPHS1, CDH1, GATA3, PDGFR-β and CD31. Protocols were modified to generate organoids to contain different compositions of specific cell types and structures, and they were tested for nephrotoxicity and kidney injury response by exposure to stimuli including cisplatin and LPS. The degree of kidney injury was assessed by various markers including KIM-1, αSMA, cPLA2, and γH2AX. The protocols enable high throughput analyses.

Results

Our novel method to generate human kidney organoids without undefined matrices yields all major cell types including proximal and distal tubules, podocytes, collecting duct, stromal cells, and endothelial cells in anatomically juxtaposed contexts. Avoidance of undefined matrices facilitates potential use of these organoids for more clinically relevant applications. Variation on the simplified and cost-effective protocols results in variation in composition of cell types and structures along with varying connectivity among cell types. Immunostaining for markers including KIM-1, LTL, αSMA, cPLA2, and γH2AX showed that kidney organoids containing multiple distinct structures present together were more sensitive to nephrotoxicants compared to organoids generated to have predominant amount of proximal tubules lacking other cell types.

Conclusion

Protocols have been developed to generate human kidney organoids without undefined matrices in a more streamlined cost effective manner with programmed relative amounts of distinct kidney component structures. Proximal tubule susceptibility to toxicity is altered by adjustments in the organoid generation that change the surrounding contextual environment of other kidney structures.

Funding

  • NIDDK Support