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Troponin release

Cor.At® cardiomyocytes provide a standardized, homogenous and reproducible cell system for the in vitro classification of a compound‘s cardio-cytotoxic potential. After incubation with test compounds, end points such as neutral red uptake can be used to determine effects which directly affect the viability and integrity of cardiac cells when compared to a non-specific reference cell type, e.g. mouse fibroblasts. Since cardiac Troponin is considered to be the gold standard biomarker for cardiac injury, e.g. after myocardial infarction, we have established a cardiac toxicity assay using Cor.At® cardiomyocytes in our standard Cor.At® Tox set-up, but with Troponin release as read-out. 

Cor.At® cardiomyocytes were thawed and cultured in 96 well plates in standard Cor.At® medium for 3 days, followed by 3 days of compound treatment. Supernatant was collected after 24h, 48h, and 72h, and amount of Troponin was quantified using the HIGH SENSITIVITY MOUSE CARDIAC TROPONIN I ELISA KIT, Life Diagnostics, Inc., Cat. No. 2010 1 HS. 

The assay detects Troponin I in a range from 0.156 to 10 ng/ml. Control values (Cor.At® cells treated with solvent only) was about 0.6 ng/mL, the highest measured value of treated cells 3.8 ng/mL. All values layed within the range of the assay's standard curve. All calculations were performed with blank-corrected values. Control OD values were 6x above blank OD values (Blank = medium only w/o cells).

The determination of cardiac troponin was established as the "gold standard" for the detection of cardiac injury [1]

Advantages troponin release analysis
  • Integrated cell culture model representing primary-like cardiac physiology.
  • Predictive, relevant and robust test system capable of medium to high throughput screening.
  • Gold standard biomarker of cardiac injury as read-out in a high-sensitivity ELISA format.
  • Comparison with possible effects on cardiac electrophysiology and mitochondrial function within the same cell system.
  • Either standardized or customized protocols available with respect to drug treatment period, concentrations, and read-out.
  • Library-specific validation studies possible.