Differentiation of cardiomyocytes and generation of human engineered heart tissue
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.
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Optimal cardiogenic mesoderm induction for all hESC and hiPSC lines was found at 1-4 μM CHIR99021 concentrations, rather than the typical 10-12 μM range reported for monolayer methods.
“Optimal cardiogenic mesoderm induction for all human embryonic stem cell (hESC) and induced pluripotent stem cell (hiPSC) lines was found at 1–4 μM CHIR99021 concentrations, rather than the typical 10–12 µM range reported for monolayer methods14,21–27.”
From Self-assembling human heart organoids for the modeling of cardiac development and congenital heart disease (2021) · cites this paper as [23]
BMP4 and Activin A induce cardiac mesoderm in the embryo, and established differentiation protocols using them differentiate to various cardiac mesoderm progenitors.
“These growth factors lead to the induction of cardiac mesoderm in the embryo70 and established differentiation protocols using them show effective differentiation to various cardiac mesoderm progenitors23,70,71.”
From Self-assembling human heart organoids for the modeling of cardiac development and congenital heart disease (2021) · cites this paper as [23]
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- Authors
- Breckwoldt, K., Letuffe-Brenière, D., Mannhardt, I., Schulze, T., Ulmer, B., Werner, T., Benzin, A., Klampe, B., Reinsch, M. C., Laufer, S., Shibamiya, A., Prondzynski, M., Mearini, G., Schade, D., Fuchs, S., Neuber, C., Krämer, E., Saleem, U., Schulze, M. L., Rodriguez, M. L., Eschenhagen, T., & Hansen, A.
- Affiliations
- University Medical Center Hamburg-Eppendorf (DE)
- TU Dortmund University (DE)
- Corresponding author
- Hansen, Arne
- ORCID
- Venue
- Nature Protocols
- Published
- 2017-05-11
- Volume
- 12
- Issue
- 6
- Pages
- 1177-1197
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W2612983729
- Cited by
- 272
- References
- 60
- Citations by year
- 2026: 23 · 2025: 33 · 2024: 26 · 2023: 20 · 2022: 35 · 2021: 41 · 2020: 40 · 2019: 28 · 2018: 18 · 2017: 8
- FWCI
- 10.49
- Type
- journal article
- Language
- en
- Update notices
- correction (2019-08-05)
- Open access
- closed
- Locations
- Nature Protocols (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Congenital heart defects research
- Keywords
- cardiomyocyte differentiation, human induced pluripotent stem cells, engineered heart tissue, cardiomyocyte contractility, embryoid bodies, cardiac differentiation
- MeSH terms
- Cell Differentiation, Humans, Tissue Engineering, Myocytes, Cardiac, Induced Pluripotent Stem Cells
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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