Figure 1.

Experimental setup and mitochondrial and glycolysis stress test parameters

(A) Experimental setup for profiling bioenergetics in intestine-derived organoids. This setup can be used to analyze organoid bioenergetics upon various treatments. Examples are (i) short treatment with metabolic inhibitors that are injected during the assay, (ii) incubation of organoids in the presence of different metabolic substrates such as glucose vs palmitate in the assay medium, (iii) induction or gene knockdown after re-plating organoids for analysis, or (iv) induction of organoid differentiation or cell-type enrichment which is already started prior to re-plating the organoids for analysis. ∗For other organoid types or specific differentiation protocols, culturing for longer than 10 days might be required (B and C). Setup of and respiratory parameters measured during (B) a mitochondrial stress test or (C) a glycolysis stress test.