Protocol to analyze bioenergetics in single human induced-pluripotent-stem-cell-derived kidney organoids using Seahorse XF96

STAR Protoc. 2023 Mar 17;4(1):101999. doi: 10.1016/j.xpro.2022.101999. Epub 2023 Jan 5.


Metabolic derangement is a key culprit in kidney pathophysiology. Organoids have emerged as a promising in vitro tool for kidney research. Here, we present a fine-tuned protocol to analyze bioenergetics in single human induced-pluripotent-stem-cell (iPSC)-derived kidney organoids using Seahorse XF96. We describe the generation of self-organized three-dimensional kidney organoids, followed by preparation of organoids for Seahorse XF96 analysis. We then detail how to carry out stress tests to determine mitochondrial and glycolytic rates in single kidney organoids.

Keywords: Cell Differentiation; Metabolism; Organoids; Stem Cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation
  • Energy Metabolism
  • Humans
  • Induced Pluripotent Stem Cells*
  • Kidney
  • Organoids
  • Solitary Kidney* / metabolism