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

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

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.

MeSH terms

  • Cell Culture Techniques / methods*
  • Cell Differentiation / drug effects
  • Cisplatin / pharmacology
  • Humans
  • Nephrons / cytology*
  • Organoids / cytology*
  • Organoids / drug effects
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / drug effects

Substances

  • Cisplatin