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Kidney Week

Abstract: FR-OR047

Generation of Functional Human Kidney Tissues from Metanephric Nephron Progenitors and Ureteric Bud Cells Separately Differentiated from Human iPS Cells

Session Information

  • Development and Stem Cells
    November 08, 2019 | Location: 152, Walter E. Washington Convention Center
    Abstract Time: 05:42 PM - 05:54 PM

Category: Development, Stem Cells, and Regenerative Medicine

  • 500 Development, Stem Cells, and Regenerative Medicine

Authors

  • Tsujimoto, Hiraku, Center for iPS Cell Research and Application, Kyoto University, Kyoto City, Kyoto, Japan
  • Araoka, Toshikazu, Center for iPS Cell Research and Application, Kyoto University, Kyoto City, Kyoto, Japan
  • Ryosaka, Makoto, Center for iPS Cell Research and Application, Kyoto University, Kyoto City, Kyoto, Japan
  • Mae, Shin-ichi, Center for iPS Cell Research and Application, Kyoto University, Kyoto City, Kyoto, Japan
  • Osafune, Kenji, Center for iPS Cell Research and Application, Kyoto University, Kyoto City, Kyoto, Japan
Background

Chronic kidney disease (CKD) affects more than 10% of the global population. The lack of effective curative options has led to research on regenerative therapies using stem cells. Accordingly, recent studies using human induced pluripotent stem cells (hiPSCs) have developed protocols to induce kidney-lineage cells and reconstruct kidney organoids. However, no reports have generated human kidney tissues by recapitulating nephrogenesis using metanephric nephron progenitors (NPs) and ureteric bud (UB) cells induced separately from hiPSCs, in which NP-derived glomeruli and renal tubules and UB-derived collecting ducts are interconnected. Furthermore, no in vivo imaging studies have directly demonstrated that hiPSC-derived kidney organoids produce urine.

Methods

We separately and efficiently induced metanephric NPs and UB cells from hiPSCs in the original 2D differentiation culture conditions, co-cultured these two progenitors using bioreactors and performed immunofluorescent analysis using the CUBIC tissue clearing method. In addition, we transplanted mixed aggregates from the two progenitors into immunodeficient mice and examined them using in vivo multiphoton microscopy.

Results

After co-culture of the two progenitors, NPs constructed SIX1(+) active nephrogenic niches close to UB tips and S-shaped body-like structures. They further organized kidney structures that contained glomeruli, proximal and distal tubules, Henle’s loops and collecting ducts in vitro and in vivo. By using two hiPSC lines that constitutively express fluorescent reporter proteins (GFP or mCherry), we demonstrated that the connecting points of GFP(+) NP-derived distal tubules and mCherry(+) UB-derived collecting ducts showed a marker expression pattern consistent with their counterparts in human embryonic kidneys, indicating that they were functionally interconnected. Furthermore, the intravenous injection of fluorescent-conjugated dextran confirmed that the hiPSC-derived glomeruli were functionally integrated with the host vasculature. Moreover, we observed urine-like dextran accumulation in the hiPSC-derived Bowman's space in vivo.

Conclusion

Our culture system should contribute to the mechanistic elucidation of human nephrogenesis and the development of regenerative therapies against kidney diseases.

Funding

  • Government Support - Non-U.S.