What is cardiotoxicity?
Cardiotoxicity or cardiac toxicity is a term used to define chemicals that are toxic to the heart, causing muscle damage or heart electrophysiology dysfunction.
The assessment of cardiotoxicity is important in the early stages of drug discovery to eliminate potentially toxic compounds from further development. This is critical to reduce the inefficiencies and high costs associated with compounds that fail during cardiac safety assessment.
Cardioactive compounds are used in clinical treatment of heart failure, arrhythmia, or other cardiac diseases. Cardiac toxicity can cause arrhythmias or heart failure. Therefore, there is a growing need for highly predictive in vitro cardiotoxicity assays that use biologically relevant 3D cell-based models and are suitable for high-throughput screening.

Mechanical movement and calcium fluxes of the cardiac spheroid can be observed from the stacks of the time-lapse acquisition images.
Evaluate cardiotoxic compound effects using human iPSC-derived cardiomyocytes
What are cardiomyocytes?
Cardiomyocytes are the cells that make up the cardiac muscles and are responsible for the heart's contractile function. Specialized cardiomyocytes like human induced pluripotent stem cell (iPSC)-derived cardiomyocytes are especially attractive cell models because they express GPCRs and ion channels that provide gene expression profiles as well as phenotypic characteristics while demonstrating spontaneous mechanical and electrical activity similar to native cardiac cells. iPSC-derived cardiomyocytes form a synchronously beating monolayer that can be used to reliably reproduce drug-associated, cardio-physiologic phenotypes using a fast, kinetic fluorescence assay that monitors changes in intracellular calcium oscillations.

In collaboration with Eurofins, we discuss the assessment of potential cardiotoxicity of compounds with CiPA initiative, the new cardiac safety testing paradigm that includes in vitro assays using hiPSC-CM.
The CiPA Initiative – Comprehensive in vitro Proarrhythmia Assay
Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) can be used to identify and evaluate the effects of potentially cardiotoxic compounds. The US FDA has sponsored an initiative to evaluate compounds in this category called the Comprehensive in vitro Proarrhythmia Assay (CiPA) initiative. There are 28 compounds that have various effects on cardiomyocytes that can be toxic. The purpose of this initiative is to determine assays that are predictive earlier in the drug discovery process to help prevent failure in either late-stage development or even after release. Several compounds on the CiPA list have been pulled from the market due to cardiac toxicity. It is possible to identify toxicity in these compounds using both measurement of calcium flux, and high content imaging. The development of highly predictive in vitro assays suitable for high-throughput screening is critical to improving the inefficiencies and high costs associated with cardiac safety compound failure.
High-throughput kinetic screening solution for toxicology and lead compound identification
The FLIPR® Penta High-Throughput Cellular Screening System is a flexible and reliable real-time kinetic high-throughput cellular assay screening system for identifying early leads against GPCR and ion channel receptors. It is designed to collect signals from all wells at the same time. The system can also monitor changes in intracellular Ca2+ flux (calcium oscillations) associated with cardiomyocyte contractions using the EarlyTox Cardiotoxicity Kit. The assays use iPSC-derived cardiomyocytes loaded with a calcium-sensitive dye and allow you to monitor the compound impact on oscillating calcium within the cells as they beat.
ScreenWorks® Peak Pro Software offers the ability to analyze multi-peak calcium oscillation responses of cells—in particular, cardiomyocyte beat rate as well as temporal characteristics of peaks including rise, decrease, and amplitude. These features are important to better understand in vitro cardiomyocyte function and the impact of toxic compounds that induce cardiac abnormalities such as hERG blocking potassium channels. As a result, you can fail or further monitor specific compounds sooner, and better prioritize the most promising, safe, leads to take forward into preclinical studies.
Spontaneous calcium oscillation acquired with FLIPR Calcium Assay Kit and the FLIPR system from iPSC cardiomyocytes or 3D cardiac spheroids can be paired with cytotoxicity and mitochondrial integrity data from the ImageXpress confocal system for comprehensive phenotypic analysis and bioactivity profiling of both pharmaceutical and environmental compounds.

The FLIPR Penta system allows you to measure and analyze peaks of human-derived induced pluripotent stem cells (hiPSCs), differentiated into cardiomyocytes and neurons, up to 100 times per second and quickly cherry pick events such as Early Afterdepolarization-like events (EAD-like events), fibrillation, and irregular beating.
Applications for characterizing cardiotoxic compounds using stem cell-derived cardiomyocytes
Traditional methods for characterizing cardiotoxic compounds are labor-intensive and slow. Manual patch clamping and automated electrophysiology are limited to the analysis of single channels on individual cells and have high costs and low throughput. Other, higher throughput methods require the exporting of data into additional, often complex, software for analysis or time-consuming manual analysis of the data.
Our application assets below demonstrate how ScreenWorks Peak Pro Software running on the FLIPR system with EarlyTox Cardiotoxicity Kit can quickly and easily characterize cardiotoxic compounds using stem cell-derived cardiomyocytes. In addition, the FLIPR system can be used to analyze neurotoxic effects upon Neuronal spheroids in much the same manner as iPSC-derived cardiomyocytes.
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3D cardiac spheroids
There is an increasing interest in exploring the use of three-dimensional (3D) spheroids for modeling tissue biology and toxicity assessment. The development of quantitative assays in higher throughput using 3D cultures is an active area of investigation. Studying the formation of 3D spheroids derived from human induced pluripotent stem cells (iPSC), using both high-content imaging (HCI) and fast kinetic fluorescence imaging helps measures the impact of various compounds on the beating rate and pattern of cardiac spheroids as monitored by changes in intracellular calcium levels with calcium-sensitive dyes.
Assessment of compound-induced pro-arrhythmic effects in vitro
Using human iPSC-derived cardiomyocytes as an in vitro model, we evaluated the responses and concentration dependence to 28 drugs linked to low, intermediate, and high torsades de pointes (TdP) risk categories. The impact of various compounds on the beating rates and patterns of cardiomyocyte spontaneous activity was monitored by changes in intracellular Ca2+ oscillations measured by fast kinetic fluorescence with calcium-sensitive dyes on the FLIPR Penta system.
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Calcium oscillation
Induced pluripotent stem cell (iPSC)- derived cardiomyocytes are a particularly attractive in vitro model system due to their use for evaluation of compound effects on both cardiac function and safety. Calcium signal oscillation reflects changes in cytoplasmic calcium concentration making it possible to use a calcium sensitive dye such as the EarlyTox™ Cardiotoxicity Kit.
Calcium oscillation in 3D neuronal spheroids
3D neural spheroids contain a neural network enriched in synapses, creating a highly functional neuronal circuitry that displays spontaneous, synchronized, readily detectable calcium oscillations.
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Cardiomyocyte assays
Stem cell-derived human cardiomyocytes have phenotypic characteristics and electrophysiological profiles similar to those of native human cardiac cells. Cardiomyocytes in culture are able to form a beating syncytium, which behaves similarly to native cardiomyocytes. Oscillation of intracellular calcium levels occurring with synchronized contractions of the cells can be monitored using a calcium-sensitive dye, and treatment-induced changes in the pattern of oscillation can be monitored via changes in fluorescent signal over time.
- Multiparametric assays for cardiomyocytes on a single platform
- Live Cell Beating Assay Using Human iPSC-derived Cardiomyocytes for Evaluation of Drug Efficacy and Toxicity
- Compound effects upon calcium transients in beating Axiogenesis Cor.4U human iPS cell-derived cardiomyocytes
- Automated Functional Cellular Analyses of Human iPS-derived Cardiomyocytes
Cardiotoxicity assays
Cardiac toxicity is responsible for a large percentage of new drugs that fail in clinical trials. The development of highly predictive in vitro assays suitable for highthroughput screening is critical to improve the ineffeciencies and high costs associated with cardiac safety compound failure.
- High throughput cardiotoxicity assays using stem cell-derived cardiomyocytes
- Predictive Assays for High Throughput Assessment of Cardiac Toxicity and Drug Safety
- EarlyTox Cardiotoxicity Kit provides biologically relevant cardiotoxicity data earlier in the drug discovery process
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eBooks
Enjoy a compilation of eBooks related to the real-time, kinetic high-throughput cellular assay screening solution, FLIPR system.
Publications on cardiotoxicity
Curated cardiotoxicity articles from Molecular Devices subject matter experts, featured in trade journals offered on pubmed.gov and Research Gate.
- Assessment of Beating Paramaters in Human Induced Pluripotent Stem Cells Enables Quantitative in Vitro Screening for Cadiotoxicity
- Phenotypic Assays for Characterizing Compound Effects on Induced Pluripotent Stem Cell-Derived Cardiac Spheroids
- In Vitro Cardiotoxicity Assessment of Environmental Chemicals Using an Organotypic Human Induced Pluripotent Stem Cell-Derived Model
- Multiparameter In Vitro Assessment of Compound Effects on Cardiomyocyte Physiology Using iPSC Cells
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Toxicology
Toxicology is the study of adverse effects of natural or man-made chemicals on living organism. It is a growing concern in our world today as we are exposed to more and more chemicals, both in our environment and in the products we use.
Latest Resources
Resources of Cardiotoxicity
Blog
3D organoids and automation of complex cell assays [Podcast]
3D organoids and automation of complex cell assays [Podcast]
As we enter the era of sophisticated drug discovery with gene therapy and personalized medicine, we need to be prepared to study complex diseases, assess the therapeutic effect of…
Publications
Q&A: What the transition away from animal testing could mean for drug discovery
Q&A: What the transition away from animal testing could mean for drug discovery
In September, the U.S. Senate unanimously passed the FDA Modernization Act 2.0, which would lift an 84-year-old federal mandate for animal testing for toxicity studies. While the bill doesn’…
News
Molecular Devices and HeartBeat.bio announce collaboration to automate and scale cardiac organoids for high-throughput screening in drug discovery
Molecular Devices and HeartBeat.bio announce collaboration to automate and scale cardiac organoids for high-throughput screening in drug discovery
SAN JOSE, Calif., and VIENNA, Sept. 12, 2022 – Molecular Devices, LLC., a leading provider of high-performance life science solutions, and HeartBeat.bio AG, a biotech company developing card…
Blog
Stem Cell Science and Regenerative Medicine – Technology and Methods Presented at ISSCR 2022
Stem Cell Science and Regenerative Medicine – Technology and Methods Presented at ISSCR 2022
It was another great year at ISSCR 2022! Leaders from across the globe came together to discuss new technologies, share insights, and explore the newest breakthroughs in stem cell…
Scientific Poster
High-throughput assessment of compound-induced pro-arrhythmic effects in human IPSC-derived cardiomyocytes
High-throughput assessment of compound-induced pro-arrhythmic effects in human IPSC-derived cardiomyocytes
Development of biologically relevant and predictive cell-based assays for compound screening and toxicity assessment is a major challenge in drug discovery. The focus of this study was to es…
Application Note
Phenotypic characterization of neuroactive compound effects
Phenotypic characterization of neuroactive compound effects
The FLIPR Penta system is powered by a high-speed EMCCD camera and the new ScreenWorks Peak Pro 2 software. The system allows measurement and analysis of complex patterns of calcium…
Application Note
Functional evaluation of neurotoxic and neuroactive compound effects in iPSC-derived 3D neural co-cultures
Functional evaluation of neurotoxic and neuroactive compound effects in iPSC-derived 3D neural co-cultures
There is increasing interest in using more complex, biologically-relevant, and predictive cell-based platforms for assay development and compound screening.
Scientific Poster
Multi-parametric assessment of compound-induced pro-arrhythmic effects in human iPSC-derived cardiomyocytes
Multi-parametric assessment of compound-induced pro-arrhythmic effects in human iPSC-derived cardiomyocytes
Development of biologically relevant and predictive cell-based assays for compound screening and toxicity assessment is a major challenge in drug discovery
eBook
Phenotypic Screening With iPSC-Derived Cardiomyocytes and Neurons
Phenotypic Screening With iPSC-Derived Cardiomyocytes and Neurons
This new eBook discusses early drug safety testing using HTS iPSC-derived cells with imaging. Download it now!
eBook
Identify Targets Like a Pro
Identify Targets Like a Pro
Our FLIPR Tetra® High-Throughput Cellular Screening System is fast, reliable, and remarkably easy to configure. The system is optimized for use with both fluorescence and luminescence, and a…
Application Note
Measure Calcium Oscillation in iCell Cardiomyocytes2 on the FLIPR Tetra System
Measure Calcium Oscillation in iCell Cardiomyocytes2 on the FLIPR Tetra System
Shorten growth timelines when screening compound effects on calcium transients in beating cardiomyocytes by using the iCell® Cardiomyocytes2 Kit (Cellular Dynamics International). iCell…
eBook
Optimized Assays for Breakthrough Research
Optimized Assays for Breakthrough Research
Biology research has been greatly simplified by the availability of standardized assay kits, instruments, and assay protocols from commercial manufacturers.
Application Note
Multiparametric assays for cardiomyocytes on a single platform
Multiparametric assays for cardiomyocytes on a single platform
Recently, it has been shown that changes in intracellular calcium flux associated with cardiomyocyte contractions can be monitored on a fluorescence imaging plate reader (FLIPR Tetra System…
Application Note
Assessment of drug effects on cardiomyocyte physiology using human iPSC-derived cardiac spheroids
Assessment of drug effects on cardiomyocyte physiology using human iPSC-derived cardiac spheroids
Cell models are becoming more complex in order to better mimic in vivo microenvironments and provide greater predictivity for compound efficacy and toxicity. There is an increasing interest…
Application Note
EarlyTox Cardiotoxicity Kit provides biologically relevant cardiotoxicity data earlier in the drug discovery process
EarlyTox Cardiotoxicity Kit provides biologically relevant cardiotoxicity data earlier in the drug discovery process
The EarlyTox Cardiotoxicity Kit contains a novel calcium sensitive dye and a proprietary masking technology optimized for measuring changes in cytoplasmic calcium which are associated with…
Application Note
High throughput cardiotoxicity assays using stem cell-derived cardiomyocytes
High throughput cardiotoxicity assays using stem cell-derived cardiomyocytes
Eliminate cardiotoxic compounds and identify potential cardiac drug candidates earlier in the drug discovery process with the FLIPR Tetra System, together with ScreenWorks Peak Pro Software…
Application Note
Compound effects upon calcium transients in beating Axiogenesis Cor.4U human iPS cell-derived cardiomyocytes
Compound effects upon calcium transients in beating Axiogenesis Cor.4U human iPS cell-derived cardiomyocytes
The assessment of cardiotoxicity is important in the early stages of drug discovery to enable elimination of potentially toxic compounds from further development. This is critical to reduce…
Scientific Poster
Phenotypic Characterization of Compound Effects on iPSC-derived Cardiac & Liver Spheroids
Phenotypic Characterization of Compound Effects on iPSC-derived Cardiac & Liver Spheroids
Explore PDF & learn the use of fast kinetic fluorescence and 3D image analysis for Phenotypic characterization of compound effects on iPSC-derived Cardiac and Liver Spheroids.
Scientific Poster
In Vitro Assessment of Drug Effects on Human iPSC-Derived Cardiac Spheroid Cultures
In Vitro Assessment of Drug Effects on Human iPSC-Derived Cardiac Spheroid Cultures
for modeling developmental and tissue biology with the goal of accelerating translational research. Such 3D models can provide different perspectives from traditional 2D cultures on the resp…
Scientific Poster
Predictive Assays for High Throughput Assessment of Cardiac Toxicity and Drug Safety
Predictive Assays for High Throughput Assessment of Cardiac Toxicity and Drug Safety
Read more about cell based assays for measuring the impact of pharmacological compounds on the rate of beating cardiomyocytes with different assay platforms.
Scientific Poster
Automated Functional Cellular Analyses of Human iPS-derived Cardiomyocytes
Automated Functional Cellular Analyses of Human iPS-derived Cardiomyocytes
This PDF explores live cell assays for measuring the impact of pharmacological compounds on rate & magnitude of beating cardiomyocytes using high content imaging & electrophysiology.
Scientific Poster
Live Cell Beating Assay Using Human iPSC-derived Cardiomyocytes for Evaluation of Drug Efficacy and Toxicity
Live Cell Beating Assay Using Human iPSC-derived Cardiomyocytes for Evaluation of Drug Efficacy and Toxicity
Learn about cell assays for measuring the impact of pharmacological compounds on rate & magnitude of beating cardiomyocytes using FLIPR Tetra & ImageXpress Micro Systems.
Publications
Phenotypic Assays for Characterizing Compound Effects on Induced Pluripotent Stem Cell-Derived Cardiac Spheroids
Phenotypic Assays for Characterizing Compound Effects on Induced Pluripotent Stem Cell-Derived Cardiac Spheroids
Development of more complex, biologically relevant, and predictive cell-based assays for compound screening is a major challenge in drug discovery. The focus of this study was to establish h…
Publications
In Vitro Cardiotoxicity Assessment of Environmental Chemicals Using an Organotypic Human Induced Pluripotent Stem Cell-Derived Model
In Vitro Cardiotoxicity Assessment of Environmental Chemicals Using an Organotypic Human Induced Pluripotent Stem Cell-Derived Model
An important target area for addressing data gaps through in vitro screening is the detection of potential cardiotoxicants. Despite the fact that current conservative estimates relate at lea…
Publications
Assessment of Beating Paramaters in Human Induced Pluripotent Stem Cells Enables Quantitative in Vitro Screening for Cadiotoxicity
Assessment of Beating Paramaters in Human Induced Pluripotent Stem Cells Enables Quantitative in Vitro Screening for Cadiotoxicity
Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes show promise for screening during early drug development. Here, we tested a hypothesis that in vitro assessment of multiple…
Publications
Multiparameter In Vitro Assessment of Compound Effects on Cardiomyocyte Physiology Using iPSC Cells
Multiparameter In Vitro Assessment of Compound Effects on Cardiomyocyte Physiology Using iPSC Cells
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…
Videos & Webinars

Toxicity studies of iPSC-derived cardiomyocytes and neuronal spheroids

CiPA initiative and validation of high-throughput methods to characterize compound effects in human iPSC-derived cardiomyocytes on the FLIPR Penta system

Validating Cardiac Ion Channels Using Automated Electrophysiology for the CiPA Paradigm

StemoniX microBrain 3D Assay Ready Plates for HTS