Functional Mitochondrial Toxicity Assay (Seahorse XFe96)

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 delivers consistent, high quality data with the flexibility to adapt protocols based on specific customer requirements.

Mitochondrial toxicity and its measurement in vitro using the Seahorse Flux Analyzer.

• Impairment of mitochondrial function is increasingly implicated in the etiology of drug-induced toxicity¹.
• The Seahorse XF^e96 extracellular flux analyzer is used to detect, in real time, effects of compounds on oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) in order to assess mitochondrial function and cellular metabolism.
• The assay uses the mitochondrial stress test to gain an insight into cellular bioenergetics and the mechanism of mitochondrial toxicity².
• In the stress test, cells are exposed sequentially to oligomycin (ATP synthase inhibitor), FCCP (protonophoric uncoupler), and rotenone and antimycin A (electron transport inhibitors). This provides information on basal respiration, proton leak, maximum respiration rate, and non-mitochondrial respiration.
• As well as mitochondrial toxicity, the Seahorse XF^e flux analyzer can be used for other applications where a shift between mitochondrial respiration and glycolysis is observed under certain pathological states (e.g., obesity, diabetes, cancer, cardiovascular disease and neurodegenerative function).

Known mitochondrial toxicants and non-toxicants were screened in the Seahorse assay.

_{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
4) Eakins J, Bauch C, Woodhouse H et al (2016) A combined in vitro approach to improve the prediction of mitochondrial toxicants. Toxicol In Vitro 34:161–170. https://doi.org/10.1016/j.tiv.2016.03.016
5) van der Stel, W., Carta, G., Eakins, J. et al. Multiparametric assessment of mitochondrial respiratory inhibition in HepG2 and RPTEC/TERT1 cells using a panel of mitochondrial targeting agrochemicals. Arch Toxicol 94, 2707–2729 (2020). https://doi.org/10.1007/s00204...}