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Abstract: SA-OR49

Expansion of Human Induced Pluripotent Stem Cell-Derived Ureteric Bud Organoids with Repeated Branching Potential

Session Information

Category: Development, Stem Cells, and Regenerative Medicine

  • 500 Development, Stem Cells, and Regenerative Medicine

Authors

  • Ryosaka, Makoto, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan
  • Mae, Shin-ichi, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan
  • Osafune, Kenji, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan
Background

The mammalian adult kidney, metanephros, develops by the reciprocal interaction between two embryonic progenitor tissues, metanephric mesenchyme and ureteric bud (UB). UB has epithelial polarity and tubular lumens, consists of two domains, the tip and trunk, and repeats branching morphogenesis. The tip cells produce both new tip and trunk cells that further differentiate into collecting ducts (CDs). Recently, we reported a stepwise protocol to induce human induced pluripotent stem cells (hiPSCs) and embryonic stem cells (hESCs) into UB-like structures through anterior intermediate mesoderm. However, the generation of hiPSC/ESC-derived UB-like tissues that show tubular lumens or sufficient branching has not been achieved.

Methods

We established a novel method to induce hiPSCs to differentiate into induced UB (iUB) organoids. We evaluated our iUB organoids using immunostaining and single cell RNA sequencing analysis.

Results

Our iUB organoids showed RET+ tip and CK19+ trunk domains, epithelial polarity, tubular lumens and developmental potential to repeat branching morphogenesis. The isolated tip regions from the iUB organoids showed repeated branching to reconstitute the iUB organoids. We also succeeded in establishing in vitro monitoring and expansion methods for tip cells that can efficiently reconstitute iUB organoids and differentiate into CD progenitors. In addition, we confirmed that our iUB organoids recapitulate in vivo development accompanied by spatiotemporal regulation of the gene network by high-resolution transcriptome analysis using single cell RNA sequencing, which showed the reciprocal expression of UB tip and trunk markers.

Conclusion

Our induction method for iUB organoids will help elucidate the developmental mechanisms of UB branching and develop a selective differentiation method for CD cells, contributing to the creation of disease models for congenital renal abnormalities.

Funding

  • Government Support - Non-U.S.