eCiphr®Cardio
Cardiac Assessment Using Microelectrode Array
- A highly purified population of cardiomyocytes, differentiated from human induced pluripotent stem (iPS) cells (Cellular Dynamics iCell® Cardiomyocytes), are used.
- The cells are a mixture of spontaneously electrically-active atrial, nodal, and ventricular-like myocytes. They possess typical human heart cell characteristics forming electrically connected syncytial layers that beat in synchrony, and exhibit expected electrophysiological and biochemical responses upon reference drug exposure.
- Viability is maintained for an extended culture periods (up to 2 weeks) allowing for acute and chronic studies.
- Microelectrode array (MEA) is one of the most sophisticated and efficacious technologies for measuring changes in spontaneously-active cells, such as cardiomyocytes and neurons.
- Cyprotex’s eCiphr®Cardio is a cell-based assay which uses MEA recording to monitor electrophysiological activity by measuring beat rate, field potential duration, amplitude and conduction velocity.
- Unlike the patch-clamp hERG assay, eCiphr®Cardio assesses changes in all major ion channels implicated in an action potential.
- This cardiac assay provides a unique in vitro system for preclinical drug discovery, cardiotoxicity assessment, disease modeling and high throughput phenotypic screening of drug candidates
The recent applications of pluripotent stem cells and their derivatives in toxicology and drug research provide new alternatives to the standard routine tests performed by the industry and offer new strategies for chemical safety assessment.
1Wobus AM and Löser P. (2011) Arch Toxicol 85(2); 79-117
Protocol
eCiphr®Cardio cell based assay protocol
| Instrument | Maestro 48-well MEA system (Axion BioSystems) |
|---|---|
| Cell Type | Human iPS cell-derived iCell® cardiomyocytes (Cellular Dynamics International) plated and allowed to fully mature and beat synchronously |
| Assay Details | Five concentrations in duplicate (dependent on customer requirements) Single time point Additional time points and washout (optional) |
| Data Delivery | Beat rate and number Field potential duration Amplitude Conduction velocity (optional) |
Data
Data from Cyprotex's eCiphr®Cardio assay
Figure 1
Raw traces for vehicle control (0.1% DMSO) and test compound (verapamil)
Red arrows point to the field potential duration (FPD, indicative of the QT interval duration). Verapamil clearly shortens FPD as compared to 0.1% DMSO (vehicle control). Note: In order to distinguish between the two traces, the voltage for verapamil is purposely shifted upward.
Raw traces for vehicle control (0.1% DMSO) and test compound (verapamil)
Red arrows point to the field potential duration (FPD, indicative of the QT interval duration). Verapamil clearly shortens FPD as compared to 0.1% DMSO (vehicle control). Note: In order to distinguish between the two traces, the voltage for verapamil is purposely shifted upward.
| Compound | Family | Number Of Beats | Number Of Beats (AC50 µM) | Fast Na+ Slope (V/s) | Fast Na+ Slope (AC50 µM) | Fast Na+ Amplitude (mV) | Fast Na+ Amplitude (AC50 µM) | Field Potential Duration (ms) | Field Potential Duration (AC50 µM) | In vivo or ex vivo Indication | Canine Purkinje Fibre Preparation (µM) | hERG Block (IC50 µM) |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Aspirin | Irreversible cyclooxygenase inhibitor | no change | >100 | no change | >100 | no change | >100 | no change | >100 | no prolongation | not reported | not reported |
| Cisapride | Serotonin 5HT4 agonist | ↓ | 0.083 | ↓ | 0.16 | ↓ | 0.16 | ↑ | 0.15 | prolongation | 0.1 | 0.03-0.1 |
| FPL64176 | L-type Ca2+ channels activator | ↓ | 0.039 | ↓ | 0.052 | ↓ | 0.053 | ↑ | 0.044 | prolongation | not reported | not reported |
| Isoproterenol | β-adrenergic receptor agonist | ↑ | 0.18 | no change | no change | no change | no change | ↓ | 0.17 | shortening | not reported | not reported |
| Nifedipine | L-type Ca2+ channel blocker | ↑ | 0.34 | no change | no change | no change | no change | ↓ | 0.12 | no prolongation | >10 | not reported |
| Quinidine | hERG K+ channel blocker/class I antiarrhythmic | ↓ | 5.8 | ↓ | 5.0 | ↓ | 5.0 | ↑ | 2.7 | prolongation | 8.5 | 1 |
| Sotalol | β-adrenergic receptor blocker | ↓ | 44 | ↓ | 41 | ↓ | 42 | ↑ | 53 | prolongation | 100 | >30 |
| Verapamil | L-type Ca2+ channel blocker | ↑ | 0.34 | ↓ | 1.0 | ↓ | 1.0 | ↓ | 0.11 | shortening | 1 | 0.125 |
Table 1
Comparison of eCiphr®Cardio, hERG channel2,3,5,6 and ex vivo/in vivo2,4,7,8,9,10 results for compounds known for their effects on cardiac function.
eCiphr®Cardio is highly reproducible and shows strong concordance with the in vivo or ex vivo cardiac effects of multiple classes of compounds. eCiphr®Cardio is a powerful cell-based assay for assessing preclinical cardiac safety and is more informative than the traditional hERG assay and conventional animal models.
Comparison of eCiphr®Cardio, hERG channel2,3,5,6 and ex vivo/in vivo2,4,7,8,9,10 results for compounds known for their effects on cardiac function.
eCiphr®Cardio is highly reproducible and shows strong concordance with the in vivo or ex vivo cardiac effects of multiple classes of compounds. eCiphr®Cardio is a powerful cell-based assay for assessing preclinical cardiac safety and is more informative than the traditional hERG assay and conventional animal models.
References
1 Wobus AM and Löser P. (2011) Arch Toxicol 85(2); 79-117
2 Peng S et al., (2010) J Pharmacol Toxicol Methods 61(3); 277-286
3 Finlayson K et al., (2004) Eur J Pharmacol 500(1-3); 129-142
4 Nattel S and Quantz MA, (1988) Cardiovasc Res 22(11); 808-817
5 Walker BD et al., (1999) Br J Pharmacol 128(2); 444-450
6 Zhang S et al., (1999) Cir Res 84; 989-998
7 Cheng HC and Incardona J, (2009) J Pharmacol Toxicol Methods 60(2); 174-184
8 Couderc JP et al., (2008) J Electrocardiol 41(6); 595-602
9 Rona G (1985) J Mol Cell Cardiol 17(4); 291-306
10 Paakkari I (2002) Toxicol Lett 127; 279-284
Learn More
Learn more about toxicology in our popular Mechanisms of Drug-Induced Toxicity guide
Close
Contact us to discuss your ADME Tox issues or request a quote
Telephone:
Europe: +44 (0)1625 505100
North America (East Coast): +1-888-297-7683
Email:
info@evotec.eu
or fill out the form below:
Close
Close
Download file
Close