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Nat Protoc. 2017 Jan;12(1):195-207. doi: 10.1038/nprot.2016.170. Epub 2016 Dec 22.

Generation of nephron progenitor cells and kidney organoids from human pluripotent stem cells.

Author information

1
Renal Division, Brigham and Women's Hospital, Boston, Massachusetts, USA.
2
Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.
3
Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.

Abstract

A variety of protocols have been developed that demonstrate the capability to differentiate human pluripotent stem cells (hPSCs) into kidney structures. Our goal was to develop a high-efficiency protocol to generate nephron progenitor cells (NPCs) and kidney organoids to facilitate applications for tissue engineering, disease modeling and chemical screening. Here, we describe a detailed protocol resulting in high-efficiency production (80-90%) of NPCs from hPSCs within 9 d of differentiation. Kidney organoids were generated from NPCs within 12 d with high reproducibility using 96-well plates suitable for chemical screening. The protocol requires skills for culturing hPSCs and careful attention to morphological changes indicative of differentiation. This kidney organoid system provides a platform for studies of human kidney development, modeling of kidney diseases, nephrotoxicity and kidney regeneration. The system provides a model for in vitro study of kidney intracellular and intercompartmental interactions using differentiated human cells in an appropriate nephron and stromal context.

PMID:
28005067
PMCID:
PMC5278902
DOI:
10.1038/nprot.2016.170
[Indexed for MEDLINE]
Free PMC Article

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