Protocol to profile the bioenergetics of organoids using Seahorse

Addressing bioenergetics is key to evaluate the impact of metabolism on the regulation of biological processes and its alteration in disease. Organoids are in vitro grown self-organizing structures derived from healthy and diseased tis-sue that recapitulate with high fidelity the tissue of origin. Bioenergetics is commonly analyzed by Seahorse XF analysis. However, its application to orga-noid studies is technically challenging. Here, we share our in-house optimized protocols to examine organoid bioenergetics in response to drugs, gene knock-down, or to characterize the metabolism of specific cell types.For complete details on the use and execution of this protocol, please refer to Ludikhuize et al. (2020).

Automated summary

Addressing bioenergetics is crucial for evaluating the impact of metabolism on the regulation of biological processes and its alteration in disease. Organoids are in vitro self-organizing structures derived from healthy and diseased tissues that recapitulate tissue of origin accurately, but applying Seahorse XF analysis to organoid studies poses technical challenges.