Our novel Cardiotox Screen assay utilizes a combination of cellular assays comprising multi-parameter phenotypic profiling techniques to assess both the functional and structural cardiotoxicity potential of novel compounds from a single plate based assay.
Cyprotex deliver consistent, high quality data with the flexibility to adapt protocols based on specific customer requirements.
A large percentage of drugs fail in clinical studies due to cardiac toxicity; thus, development of sensitive in vitro assays that can evaluate potential adverse effects on cardiomyocytes is extremely important for drug development.
1 Sirenko O et al, (2012) J Biomol Screen 18(1); 39-53
|Cell Line||Induced pluripotent stem cell (iPSC) derived cardiomyocytes|
|Analysis Platform||Cytation 3 Cell Imaging Multi-Mode reader and Cellomics ArrayScan® XTI or CX7 (Thermo Scientific)|
|Test Compound Concentrations||8 point dose response curve with top concentration based on 100x Cmax or solubility limit. 3 replicates per concentration*|
|Compound Requirements||150 μL of a DMSO* solution to achieve 200 x top concentration maintained at 0.5% DMSO or equivalent amount in solid compound|
|Time Points||Acute and 24 hr pre-incubation*|
|Quality Controls||Negative control: 0.5% DMSO (vehicle)*
Positive control: 2 appropriate compounds
|Data Delivery||Minimum effective concentration (MEC) and AC50 value for each measured parameter; peak count, amplitude, frequency, full peak width, full width half maximum (FWHM), full rise time, rise time from 10%, full decay time, decay time to 10%, peak width at 10%, peak spacing (below 10%), cell count, nuclear size, DNA structure (DNA), calcium homeostasis (Ca2+) mitochondrial mass (Mito Mass), mitochondrial membrane potential (MMP) and cellular ATP content (ATP)*|
* other options available on request.
1 Sirenko O et al. (2012) Multiparameter in vitro assessment of compound effects on cardiomyocyte physiology using iPSC cells. J Biomol Screen 18(1): 39-53
2 Pointon A et al (2013) Phenotypic profiling of structural cardiotoxins in vitro reveals dependency on multiple mechanisms of toxicity. Toxicol Sci 132(2): 317-326
3 Laverty HG et al. (2011) How can we improve our understanding of cardiovascular safety liabilities to develop safer medicines? Br J Pharmacol 163(4): 675-693
4 Cross MJ et al. (2015) Physiological, pharmacological and toxicological considerations of drug-induced structural cardiac injury. Br J Pharmacol 172(4): 957-974
5 Ravenscroft S et al. (2016) Cardiac non-myocyte cells show enhanced pharmacological function suggestive of contractile maturity in stem cell derived cardiomyocyte microtissues. Toxicol Sci 152(1): 99-112