The generation of kidney organoids by differentiation of human pluripotent cells to ureteric bud progenitor–like cells

Abstract

This protocol presents recently developed methodologies for the differentiation of human pluripotent stem cells (hPSCs) into ureteric bud (UB) progenitor–like cells. Differentiation of human PSCs to UB progenitor–like cells allows for the generation of chimeric kidney cultures in which the human cells can self-assemble into chimeric 3D structures in combination with embryonic mouse kidney cells over a period of 18 d. UB progenitor–like cells are generated by a two-step process that combines in vitro commitment of human PSCs, whether embryonic stem cells (ESCs) or induced PSCs (iPSCs), under chemically defined culture conditions, with ex vivo cultures for the induction of 3D organogenesis. The models described here provide new opportunities for investigating human kidney development, modeling disease, evaluating regenerative medicine strategies, as well as for toxicology studies.

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Figure 1: Schematic representation of the differentiation procedure.
Figure 2: Overview of the reaggregation assay.
Figure 3: E12.5 mouse kidney aggregates formed by different starting cell numbers.
Figure 4: Morphology of mouse and chimeric aggregate cultures.
Figure 5: Embryonic renal marker expression of chimeric aggregates.
Figure 6: Adult renal marker expression in aggregate culture.

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Acknowledgements

We thank M. Schwarz for administrative support. We thank J. Kasuboski from the Waitt Advanced Biophotonics Core at the Salk Institute for Biological Studies for help with imaging processing. Y.X. was partially supported by the California Institute for Regenerative Medicine (CIRM) through a CIRM Training grant. I.S.-M. was partially supported by a Nomis Foundation postdoctoral fellowship. Work in the laboratory of J.C.I.B. was supported by grants from the G. Harold and Leila Y. Mathers Charitable Foundation and The Leona M. and Harry B. Helmsley Charitable Trust (2012-PG-MED002).

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Y.X., I.S.-M., E.N., C.R.E., J.M.C. and J.C.I.B. designed all experiments and developed the methodologies presented here. Y.X., I.S.-M., E.N. and J.C.I.B. wrote the manuscript.

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Correspondence to Juan Carlos Izpisua Belmonte.

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The authors declare no competing financial interests.

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Xia, Y., Sancho-Martinez, I., Nivet, E. et al. The generation of kidney organoids by differentiation of human pluripotent cells to ureteric bud progenitor–like cells. Nat Protoc 9, 2693–2704 (2014). https://doi.org/10.1038/nprot.2014.182

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