Mechanistic Toxicity

Cyprotex offer a number of different assays to elucidate specific mechanisms of toxicity.

These include:

Cell stress panel - assesses 36 biomarkers that represent cellular stress signalling pathways, organelle health and cellular cytotoxicity.

Apoptosis and necrosis assay – uses flow cytometry to assess mechanism of cell death through apoptosis and necrosis.

Phospholipidosis - lysosomal storage disorder characterized by excessive accumulation of intracellular phospholipids in tissues.

Steatosis - hepatic lipid processing disorder, leading to accumulation of triglycerides within the liver cells.

Lysosomal trapping – investigates accumulation of the drug in the lysosome.

Hemolysis - evaluates hemoglobin release in the plasma (as an indicator of red blood cell lysis).

Mitochondrial toxicity (glu/gal) – compares cytotoxicity using cells grown in glucose and galactose media to determine mitochondrial impairment.

Functional mitochondrial toxicity  – uses the Seahorse XF^e96 extracellular flux analyzer to determine oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) to assess cellular bioenergetics and potential mechanisms of mitochondrial toxicity.

Mitochondrial respiratory complex assay (using permeabilized cells)  – uses the Seahorse XF^e96 extracellular flux analyzer to decipher which complex is involved in the mitochondrial toxicity.

Reactive metabolite assessment – uses glutathione trapping studies with human liver microsomes and LC-MS/MS detection.

Cell viability - general cell viability assessment using single endpoints such as LDH (lactate dehydrogenase), neutral red or MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide).

Toxicological gene regulation – using qRT-PCR to understand toxicologically relevant changes in gene expression.

Custom toxicology – flexible service where we can investigate different cell lines, endpoints and time points to address particular customer issues.