Human iPSC-Derived Cardiomyocyte Networks on Multiwell Micro-electrode Arrays for Recurrent Action Potential Recordings
July 15th, 2019
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This article contains a set of protocols for the development of human induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM) networks cultured on multiwell MEA plates to reversibly electroporate the cell membrane for action potential measurements. High-throughput recordings are obtained from the same cell sites repeatedly over days.
Overview
This article presents protocols for developing human induced pluripotent stem cell-derived cardiomyocyte networks on multiwell MEA plates. The methods include reversible electroporation for action potential measurements, enabling high-throughput recordings over multiple days.
Key Study Components
Area of Science
- Neuroscience
- Cardiovascular Biology
- Electrophysiology
Background
- Human induced pluripotent stem cells (hiPSCs) can differentiate into cardiomyocytes.
- Multiwell MEA plates allow for simultaneous recording of electrical activity from multiple cell sites.
- Electroporation is a technique used to manipulate cell membranes for experimental purposes.
- Action potential measurements are crucial for assessing cardiomyocyte function.
Purpose of Study
- To establish a reliable protocol for culturing hiPSC-derived cardiomyocytes.
- To facilitate action potential measurements using multiwell MEA technology.
- To provide a framework for testing compounds in electrophysiological studies.
Methods Used
- Thawing and plating hiPSC-derived cardiomyocytes on MEA plates.
- Implementing a custom MATLAB workflow for data analysis.
- Performing quality checks on electrical activity post-plating.
- Using electroporation to induce action potentials for measurement.
Main Results
- Successful establishment of cardiomyocyte networks with spontaneous beating.
- High-quality action potential recordings obtained from multiple sites.
- Robust data processing methods developed for analyzing electrophysiological signals.
- Demonstrated viability of cardiomyocytes in multiwell MEA culture.
Conclusions
- The protocols provide a reliable method for studying cardiomyocyte electrophysiology.
- High-throughput capabilities enable extensive compound testing.
- Future applications may include drug screening and disease modeling.
Frequently Asked Questions
What are hiPSC-derived cardiomyocytes?
They are cardiomyocytes derived from human induced pluripotent stem cells, capable of differentiating into heart cells.
How does electroporation work in this context?
Electroporation temporarily permeabilizes the cell membrane to allow for the introduction of substances for measurement.
What is the significance of action potential measurements?
They provide insights into the electrical activity and health of cardiomyocytes, crucial for understanding heart function.
What challenges are associated with multiwell MEA plates?
Key challenges include proper cell plating and maintaining the quality of the MEA surfaces.
How can the data be analyzed?
Data can be analyzed using a custom MATLAB workflow designed for processing electrophysiological signals.
What applications can arise from this research?
Potential applications include drug testing, disease modeling, and understanding cardiac physiology.
Executive Industry Relevance
High-throughput, recurrent action potential recordings from hiPSC-derived cardiomyocyte networks on multiwell MEAs address a critical bottleneck in cardiac safety screening for drug discovery. This platform enables robust, quantitative electrophysiological assessment of compound effects, supporting predictive confidence and early de-risking of cardiotoxicity liabilities. The approach enhances portfolio triage by providing scalable, reproducible data for cross-program decision-making.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables interrogation of compound-induced electrophysiological changes in human-relevant cardiomyocyte networks.
- Supports mechanistic de-risking by quantifying action potential duration and morphology linked to arrhythmia risk.
- Facilitates functional target validation through repeated, longitudinal measurements in the same preparation.
Screening & Assay Development
- Provides a standardized, high-throughput assay format for reliable compound evaluation across multiple wells and time points.
- Delivers quantitative, reproducible action potential metrics for assay comparability and screening readiness.
- Enables rapid data extraction and analysis via custom MATLAB workflows, supporting scalable screening operations.
Translational & Preclinical Research
- Aligns in vitro electrophysiological readouts with translational biomarkers of cardiac risk.
- Supports continuity from early discovery through preclinical safety assessment by enabling chronic dose effect studies.
- Facilitates risk-adjusted advancement decisions based on human cell-derived predictive data.
Pipeline & Workflow Integration
This method integrates into the discovery-to-preclinical continuum, bridging early compound screening with translational cardiac safety evaluation.
- Discovery Biology: Supports hypothesis testing and mechanistic clarification of compound effects on cardiac electrophysiology.
- Screening: Provides assay readiness and reproducibility for high-throughput compound profiling.
- Analytics: Enables extraction of quantitative action potential parameters for robust statistical comparison.
- Translational Research: Connects in vitro findings to preclinical safety endpoints relevant for regulatory submission.
- Enterprise Reuse: Offers a reusable, scalable platform for ongoing cardiac safety and mechanistic studies across programs.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence in cardiac safety by leveraging human-relevant, quantitative electrophysiological data.
- Operational Value: Standardizes and streamlines high-throughput data acquisition and analysis workflows.
- Strategic Value: Improves go/no-go decisions and reduces late-stage attrition due to unforeseen cardiotoxicity.
- Portfolio Impact: Enables risk-adjusted prioritization and advancement of compounds with favorable cardiac profiles.
Implementation Considerations
- Requires expertise in hiPSC-cardiomyocyte culture and electrophysiological assay setup.
- Needs access to multiwell MEA instrumentation and custom data analysis software.
- Demands rigorous cross-team standardization for plating density and signal quality thresholds.
- Adaptation may be needed for different hiPSC lines or compound classes.
- Culture viability and electrode coverage are critical for reproducible outputs.
Biopharma FAQs
Why does null hypothesis testing matter for action potential duration analysis?
Null hypothesis testing enables objective assessment of whether observed changes in action potential duration after compound treatment are statistically significant, supporting robust target validation and de-risking in cardiac safety studies.
How does independent variable isolation fit in MEA-based drug screening?
Isolating variables such as compound concentration or exposure time in multiwell MEA assays allows precise attribution of electrophysiological effects, strengthening mechanistic insights and supporting confident decision-making in early discovery.
What do quantitative dependent variable measurements enable in this workflow?
Quantitative measurements of action potential parameters provide reproducible, scalable endpoints for comparing compound effects, enabling high-throughput screening and facilitating cross-program data integration.
Why are replication requirements critical for cross-functional collaboration?
Replication of action potential recordings across wells and time points ensures data reliability, enabling cross-functional teams to trust results for downstream safety, pharmacology, and translational research decisions.
What statistical analysis capabilities are required before implementation?
Robust statistical tools are needed to process large electrophysiological datasets, extract waveform parameters, and compare conditions, ensuring that only compounds with statistically validated safety profiles advance in the pipeline.
Chapters in this video
0:04
Title
0:56
Pre-plating of the Cryopreserved hiPSC-CM for Maturation
1:35
Multiwell MEA Plate Sterilization and Coating
2:57
hiPSC-CM Dissociation and Plating on Multiwell MEA Plate
4:09
hiPSC-CM Electroporation and Signal Acquisition
5:06
Data Segmentation and Analysis
6:40
Results: Action Potential Recordings of hiPSC-CMs
8:21
Conclusion
