An optimized protocol for metabolic measurement in 3D tumor spheroids derived from primary and established glioblastoma cells
Fig 2
Seahorse analyses showing a comparison between metabolic values in the different glioblastoma cell lines LN229, U138 and the primary cells GBM#4, GBM#10. Representative mitochondrial respiration measurement using the Seahorse XF Cell Mito Stress Test for U138, LN229, GBM#4 and GBM#10 spheroids generated from 30,000 cells. Oligomycin, FCCP, rotenone plus antimycin A are used as pharmacological tools to study mitochondrial respiration. Oligomycin inhibits ATP synthase, reducing oxygen consumption. Rotenone and antimycin A block the electron transport chain, preventing mitochondrial respiration and allowing measurement of non-mitochondrial oxygen consumption. Oligomycin reduces the oxygen consumption rate (OCR) through inhibition of ATP synthase. FCCP uncouples the proton gradient, maximizing oxygen consumption. Rotenone and antimycin A are electron transport chain inhibitors.