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Updated: May 16, 2026
Electrophysiological Analysis of human Pluripotent Stem Cell-derived Cardiomyocytes (hPSC-CMs) Using Multi-electrode Arrays (MEAs)
Published on: May 12, 2017
Micro-arrayed human embryonic stem cells-derived cardiomyocytes for in vitro functional assay
Elena Serena1, Elisa Cimetta, Susi Zatti
1Industrial Engineering Department, University of Padova, Padova, Italy.
Insights
This study developed a novel in vitro assay using human cardiomyocytes (hCMs) and micro-technologies to assess cardiac cell viability and function. The assay effectively models heart physiology for drug development and disease research.
Area of Science:
- Biomedical Engineering
- Cardiology Research
- In Vitro Assays
Background:
- The human heart exhibits limited regenerative capacity, making in vitro models crucial for cardiology research.
- Developing functional in vitro cardiac tissue is vital for pharmacological and physio-pathological studies.
- Existing models require enhancement for comprehensive analysis of cardiomyocyte viability and function.
Purpose of the Study:
- To create an advanced in vitro assay for human cardiomyocytes (hCMs).
- To enable simultaneous assessment of hCM viability and functionality.
- To support applications in drug screening and disease modeling for cardiac conditions.
Main Methods:
- Human cardiomyocytes (hCMs) were cultured on tunable poly-acrylamide hydrogels.
- Micropatterning techniques organized hCMs into a 20x20 array for controlled study.
- Characterization involved immunofluorescence, GAP-FRAP, live/dead assays, and monitoring excitation-contraction coupling.
Main Results:
- Micropatterned hCMs retained key cardiac markers and functional properties, with measurable contraction frequencies.
- The assay demonstrated sensitivity to oxidative stress (H2O2), showing suppressed contractility before affecting viability.
- A microfluidic platform was developed for localized, multi-parametric analysis within the cell array.
Conclusions:
- The developed system serves as a valuable tool for in vitro testing of human heart physiology.
- It facilitates the evaluation of multiple conditions on a representative cardiac cell model.
- This technology holds potential for accelerating therapeutic strategies and drug development in cardiology.
Introduction:
The heart is one of the least regenerative organs in the body and any major insult can result in a significant loss of heart cells. The development of an in vitro-based cardiac tissue could be of paramount importance for many aspects of the cardiology research. In this context, we developed an in vitro assay based on human cardiomyocytes (hCMs) and ad hoc micro-technologies, suitable for several applications: from pharmacological analysis to physio-phatological studies on transplantable hCMs. We focused on the development of an assay able to analyze not only hCMs viability, but also their functionality.
Methods:
hCMs were cultured onto a poly-acrylamide hydrogel with tunable tissue-like mechanical properties and organized through micropatterning in a 20×20 array. Arrayed hCMs were characterized by immunofluorescence, GAP-FRAP analyses and live and dead assay. Their functionality was evaluated monitoring the excitation-contraction coupling.
Results:
Micropatterned hCMs maintained the expression of the major cardiac markers (cTnT, cTnI, Cx43, Nkx2.5, α-actinin) and functional properties. The spontaneous contraction frequency was (0.83±0.2) Hz, while exogenous electrical stimulation lead to an increase up to 2 Hz. As proof of concept that our device can be used for screening the effects of pathological conditions, hCMs were exposed to increasing levels of H(2)O(2). Remarkably, hCMs viability was not compromised with exposure to 0.1 mM H(2)O(2), but hCMs contractility was dramatically suppressed. As proof of concept, we also developed a microfluidic platform to selectively treat areas of the cell array, in the perspective of performing multi-parametric assay.
Conclusions:
Such system could be a useful tool for testing the effects of multiple conditions on an in vitro cell model representative of human heart physiology, thus potentially helping the processes of therapy and drug development.
