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Abstract: PO0508

Human Primary Renal Tubuloids as Tools for Pathophysiology and Nephrotoxicity Assessment

Session Information

Category: Bioengineering

  • 300 Bioengineering

Authors

  • Sako, Keisuke, Brigham and Women's Hospital, Boston, Massachusetts, United States
  • Mori, Yutaro, Brigham and Women's Hospital, Boston, Massachusetts, United States
  • Ichimura, Takaharu, Brigham and Women's Hospital, Boston, Massachusetts, United States
  • Bonventre, Joseph V., Brigham and Women's Hospital, Boston, Massachusetts, United States
Background

Kidney organoids derived from human induced pluripotent stem cells can be used to study pathophysiology and nephrotoxicity using human kidney tissue. We have developed an efficient alternative way to generate homogeneous epithelial-like structures from kidney tissue derived from patients.

Methods

Human primary renal tubular epithelial cell (hRTEC) cultures were generated using non-tumor cells from partially resected human kidneys for renal cancer. Primary cells were cultured on ultra-low attachment plates for several days. The cells were then transferred into media containing matrigel, hepatocyte growth factor, fibroblast growth factor-2 and 5% fetal bovine serum. Tubuloids were treated with multiple nephrotoxicants. Morphological changes and KIM-1 expression were evaluated. Functional assays to characterize permeability and selective cellular absorption by tubuloids were conducted using fluorescent inulin and albumin. Co-culture experiments of tubuloids with bEnd.3 endothelial cells were performed.

Results

hRTEC tubuloids were generated based on the method above. We have generated a library of hRTECs derived from 25 patients. Tubuloids had polarized expression of cell surface markers, LTL, KIM-1 (apical) and Na-K-ATPase (basolateral). It took only a week to establish hRTEC cultures from patient kidneys and 2-4 weeks to form tubuloids from hRTECs. Cisplatin and palmitate-bound albumin altered the 3D structure of tubuloids. Treatment with aristolochic acid increased KIM-1 expression in a dose-dependent manner. Functional assays revealed that tubuloids reabsorb albumin from the apical side and release it into basolateral side, while being impermeable to inulin. Coculture of tubuloids and bEnd.3 cells resulted in formation of 3D capillary-like structure of bEnd.3 cells around tubuloids.

Conclusion

We generated renal tubuloids using hRTECs derived from patients. This strategy can recapitulate pathophysiology, enable nephrotoxicity screening of human kidney tissue and offers an additional approach to personalized kidney medicine.

Funding

  • NIDDK Support