期刊
NATURE PROTOCOLS
卷 12, 期 1, 页码 -出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2016.170
关键词
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资金
- National Institutes of Health [R37 DK039773, R01 DK072381]
- ReproCell Stem Cell Research grant
- Brigham and Women's Hospital Research Excellence Award
- Brigham and Women's Hospital Faculty Career Development Award
- Harvard Stem Cell Institute Seed grant
- NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES [R01DK072381, R37DK039773, R01DK039773] Funding Source: NIH RePORTER
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.
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