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

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 or-ganoids, followed by preparation of organoids for Seahorse XF96 analysis. We then detail how to carry out stress tests to determine mitochondrial and glyco-lytic rates in single kidney organoids.

Automated summary

Metabolic derangement plays a crucial role in kidney pathophysiology, and organoids have shown great promise as an in vitro tool for kidney research. In this study, we present a refined protocol for analyzing bioenergetics in single kidney organoids derived from human induced pluripotent stem cells (iPSCs) using Seahorse XF96. Our protocol includes generating self-organized three-dimensional kidney organoids and preparing them for Seahorse XF96 analysis, followed by stress tests to determine mitochondrial and glycolytic rates in the organoids.