Understand cellular bioenergetics and specific mechanisms of mitochondrial toxicity using Cyprotex’s functional mitochondrial toxicity assay.
Cyprotex’s functional mitochondrial toxicity service is a cell based assay which uses the Seahorse XFe extracellular flux analyzer is in Cyprotex's portfolio of in vitro toxicology services for measuring potential mitochondrial toxicity. Cyprotex deliver consistent, high quality data with the flexibility to adapt protocols based on specific customer requirements.
Drug-induced mitochondrial toxicity is rapidly gaining recognition within the pharmaceutical industry as a contributor to compound attrition and post-market drug withdrawals.
3 Nadanaciva S and Will Y (2011) Current Pharmaceutical Design 17; 2100-2112
|Media Assessed||Unbuffered DMEM containing 10 mM glucose, 1 mM pyruvate and 2 mM glutamine|
|Cell Types Available*||H9c2, Huh7, HepG2, MCF-7, cropreserved human hepatocytes, cryopreserved rat hepatocytes (other custom cell lines available on request)|
|Test Article Concentration||7 point dose response|
|Quality Controls||Vehicle control
Rotenone (positive control)
|Test Article Requirements||50 µL of 50 mM DMSO solution or equivalent amount of solid compound.|
|Analysis Method||Use of solid state fluorescent sensors to measure oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). Measured using the XFe96 flux analyzer (Seahorse Biosciences Inc)|
|Data Delivery||Summary report
AC50 for OCR, reserve capacity and ECAR
Minimum effective concentration (MEC) for OCR, reserve capacity and ECAR
|HCS based mitochondrial toxicity assessment
Glucose/galactose mitochondrial toxicity assessment
Known mitochondrial toxicants and non-toxicants were screened in the Seahorse assay.
|Oxygen Consumption Rate (OCR)||Reserve Capacity||Extracellular Acidification Rate (ECAR)|
|Compound||Mechanism||MEC (µM)||AC50 (µM)||MEC (µM)||AC50 (µM)||MEC (µM)||AC50 (µM)|
|Rotenone||Complex I inhibitor||0.008||0.017↓||0.01||0.021↓||0.01||0.016↑|
|2-Thenoyltrifluoroacetone||Complex II inhibitor||6.5||46.4↓||5||17.5↓||48||35.8↑|
|Myxothiazol||Complex III inhibitor||0.1||0.18↓||3||1.8↓||3||1.0↑|
|Antimycin A||Complex III inhibitor||0.01||0.012↓||0.01||0.008↓||0.01||0.010↑|
|Oligomycin||Complex V inhibitor (ATP-synthase inhibitor)||0.1||0.11↓||NR||NR||0.3||0.12↑|
|Carbonyl cyanide 3-chlorophenylhydrazone (CCCP)||Uncoupler||0.1||0.25↑||10||1.7↓||0.1||0.10↑|
|Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP)||Uncoupler||0.1||0.14↑||1||1.0↓||0.1||0.044↑|
|UK-5099||Pyruvate transport inhibitor||19.3||92.1↓||0.1||2.3↓||0.09||NR|
1 Dykens JA and Will Y (2007) The significance of mitochondrial toxicity testing in drug development. Drug Discovery Today 12; 777-785
2 Brand MD and Nicholls DG (2011) Assessing mitochondrial dysfunction in cells. Biochem J 435; 297–312
3 Nadanaciva S and Will Y (2011) New insights in drug-induced mitochondrial toxicity. Current Pharmaceutical Design 17; 2100-2112
Learn more about toxicology in our popular Mechanisms of Drug-Induced Toxicity guide
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