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Abstract: SA-PO003

Tubuloids on Primary Human Renal Tubular Epithelial Cells (hRTECs) Recapitulates Cisplatin-Induced Kidney Injury

Session Information

Category: Bioengineering

  • 400 Bioengineering

Authors

  • Nakao, Yuki, Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan
  • Mori, Yutaro, Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan
  • Mori, Makiko, Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan
  • Mandai, Shintaro, Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan
  • Fujiki, Tamami, Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan
  • Ando, Fumiaki, Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan
  • Susa, Koichiro, Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan
  • Mori, Takayasu, Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan
  • Sohara, Eisei, Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan
  • Uchida, Shinichi, Department of Nephrology, Tokyo Medical and Dental University, Tokyo, Japan
Background

Kidney organoids derived from human pluripotent stem cells has been an attracting pathophysiological model recapitulating a response of human kidney to drugs in recent years. Here, we have developed an alternative way to make more homogeneous epithelial-like structures called tubuloids based on primary human renal tubular epithelial cells (hRTECs) and tested their efficacy by administering cisplatin.

Methods

hRTECs were obtained from the non-tumor kidney tissue removed from patients with renal or ureteric malignancies. Renal cortex was diced and digested with collagenase. Tubules were seeded on plates with serum-free media containing epidermal growth factor. After passage, cells were cultured on ultra-low attachment plates for several days and transferred into media containing matrigel, epithelial growth factor, hepatocyte growth factor, fibroblast growth factor-2 and 5% fetal bovine serum. After tubuloids were completed, cisplatin was administered at a concentration of 0.2 – 20.0 μg/ml.

Results

Tubuloids expressed LTL and Megalin, indicating that they were highly differentiated structures composed of proximal tubular epithelial cells. Treatment of tubuloids with cisplatin increased γH2AX, a marker for DNA damage, in a dose-dependent manner. KIM-1, a marker of kidney injury, and cleaved caspase-3, a marker for apoptotic signals were expressed as well. Vimentin, an intermediate filament, was also upregulated by cisplatin treatment, suggesting that tubular epithelial cells were in the process of epithelial-mesenchymal transition. Enhancement of the NF-κB and IL-1β was also observed. These findings may recapitulate the events of post-acute kidney injury, which, after an acute phase response, acquires senescence-associated secretory phenotype (SASP) and induces inflammation and fibrosis.

Conclusion

We succeeded in establishing a model of cisplatin-induced kidney injury based on tubuloids using hRTECs. Tubuloids can be utilized to simulate the response of epithelial cells to toxins and therapeutic agents. This alternative way is potentially an excellent tool not only as cisplatin-induced kidney injury, but also as a pathological model for various renal diseases including chronic kidney disease.

Funding

  • Commercial Support – Bayer Yakuhin, Ltd.