Related Experiment Video
Updated: Mar 2, 2026
Construction of Defined Human Engineered Cardiac Tissues to Study Mechanisms of Cardiac Cell Therapy
Published on: March 1, 2016
Differentiation of cardiomyocytes and generation of human engineered heart tissue
Kaja Breckwoldt1, David Letuffe-Brenière1, Ingra Mannhardt1
1Department of Experimental Pharmacology and Toxicology, Cardiovascular Research Center, University Medical Center Hamburg-Eppendorf, and DZHK (German Center for Cardiovascular Research), Partner Site Hamburg/Kiel/Lübeck, Hamburg, Germany.
Insights
This study presents a protocol for generating engineered heart tissues from human induced pluripotent stem cells (hiPSCs) to measure cardiomyocyte contractility. This method overcomes limitations of single-cell assays for studying cardiac function.
Area of Science:
- Stem Cell Biology
- Cardiovascular Research
- Tissue Engineering
Background:
- Human induced pluripotent stem cells (hiPSCs) are crucial for disease modeling and drug discovery.
- Current methods using hiPSC-derived cardiomyocytes (hiPSC-CMs) in single-cell assays limit the assessment of contractile force due to random cell orientation.
- There is a need for robust protocols to evaluate the functional properties of hiPSC-CMs in a more physiologically relevant context.
Purpose of the Study:
- To describe a standardized protocol for differentiating hiPSCs into cardiomyocytes.
- To detail the generation of fibrin-based engineered heart tissues (EHTs) in a strip format from hiPSC-CMs.
- To enable the measurement of contractile force and other physiological parameters of hiPSC-CMs under auxotonic stretch conditions.
Main Methods:
- Parallel expansion of hiPSCs and standardized generation of defined embryoid bodies.
- Growth factor and small-molecule-based cardiac differentiation of hiPSCs into cardiomyocytes within 14 days.
- 3D assembly of cardiomyocytes into fibrin-based EHTs for contractility measurements 10-15 days post-casting.
Main Results:
- The protocol successfully differentiates hiPSCs into cardiomyocytes.
- Fibrin-based EHTs in a strip format are generated, allowing for auxotonic stretch.
- Contractility measurements can be performed on EHTs, providing functional data on hiPSC-CMs.
Conclusions:
- This protocol provides a standardized method for generating functional engineered heart tissues from hiPSCs.
- The developed EHTs enable the characterization of cardiomyocyte contractility, overcoming limitations of single-cell assays.
- This approach facilitates the recapitulation of adult human cardiomyocyte properties for research and therapeutic applications.
Abstract:
Since the advent of the generation of human induced pluripotent stem cells (hiPSCs), numerous protocols have been developed to differentiate hiPSCs into cardiomyocytes and then subsequently assess their ability to recapitulate the properties of adult human cardiomyocytes. However, hiPSC-derived cardiomyocytes (hiPSC-CMs) are often assessed in single-cell assays. A shortcoming of these assays is the limited ability to characterize the physiological parameters of cardiomyocytes, such as contractile force, due to random orientations. This protocol describes the differentiation of cardiomyocytes from hiPSCs, which occurs within 14 d. After casting, cardiomyocytes undergo 3D assembly. This produces fibrin-based engineered heart tissues (EHTs)-in a strip format-that generate force under auxotonic stretch conditions. 10-15 d after casting, the EHTs can be used for contractility measurements. This protocol describes parallel expansion of hiPSCs; standardized generation of defined embryoid bodies, growth factor and small-molecule-based cardiac differentiation; and standardized generation of EHTs. To carry out the protocol, experience in advanced cell culture techniques is required.
