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Induced pluripotent stem cell-derived cardiomyocyte in vitro models: benchmarking progress and ongoing challenges

Nature Methods volume 22, pages 24–40 (2025)Cite this article

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Abstract

Recent innovations in differentiating cardiomyocytes from human induced pluripotent stem cells (hiPSCs) have unlocked a viable path to creating in vitro cardiac models. Currently, hiPSC-derived cardiomyocytes (hiPSC-CMs) remain immature, leading many in the field to explore approaches to enhance cell and tissue maturation. Here, we systematically analyzed 300 studies using hiPSC-CM models to determine common fabrication, maturation and assessment techniques used to evaluate cardiomyocyte functionality and maturity and compiled the data into an open-access database. Based on this analysis, we present the diversity of, and current trends in, in vitro models and highlight the most common and promising practices for functional assessments. We further analyzed outputs spanning structural maturity, contractile function, electrophysiology and gene expression and note field-wide improvements over time. Finally, we discuss opportunities to collectively pursue the shared goal of hiPSC-CM model development, maturation and assessment that we believe are critical for engineering mature cardiac tissue.

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Fig. 1: hiPSC culture and differentiation.
Fig. 2: Variability in hiPSC-CM platforms.
Fig. 3: Maturation techniques used in hiPSC-CM culture.
Fig. 4: Methods of assessing hiPSC-CM functionality and maturation.
Fig. 5: Quantification of hiPSC-CM functionality and maturation.

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Data availability

The data supporting the findings of this field-wide analysis are available within the paper and its Supplementary Information. Additionally, the dataset has been deposited at Dryad and will continue to be updated as new manuscripts are published in this area of research (https://doi.org/10.5061/dryad.ksn02v7bh)34.

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Acknowledgements

This work was funded by the National Science Foundation (NSF) Engineering Research Center on Cellular Metamaterials (EEC-1647837). J.K.E acknowledges financial support from the National Institutes of Health National Heart, Lung, and Blood Institute (F31 HL158195). S.J.D. and B.M.B. acknowledge financial support from the NSF (2033654). S.J.D. acknowledges support from the National Institutes of Health (T32-DE007057 and T32-HL125242). J.K.E. and X.G. acknowledge funding support from the NSF Graduate Research Fellowship Program. L.L. acknowledges support provided by the Florida Heart Research Foundation. C.S.C. acknowledges support from the NSF (CMMI-1548571 and DGE-2244366) and the Paul G. Allen Frontiers Group Allen Distinguished Investigator Program.

Author information

Author notes

  1. These authors contributed equally: Jourdan K. Ewoldt, Samuel J. DePalma.

Authors and Affiliations

  1. Department of Biomedical Engineering, Boston University, Boston, MA, USA

    Jourdan K. Ewoldt, M. Çağatay Karakan, Xining Gao, Micheal A. McLellan, Alice E. White & Christopher S. Chen

  2. Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA

    Samuel J. DePalma, Maggie E. Jewett & Brendon M. Baker

  3. Department of Mechanical Engineering, Boston University, Boston, MA, USA

    M. Çağatay Karakan, Paria Mir Hashemian, Marshall Ma, Thomas G. Bifano, Alice E. White & Emma Lejeune

  4. Photonics Center, Boston University, Boston, MA, USA

    M. Çağatay Karakan, Paria Mir Hashemian, Marshall Ma, Thomas G. Bifano & Alice E. White

  5. Department of Biomedical Engineering, Florida International University, Miami, FL, USA

    Yih-Mei Lin & Sharan Ramaswamy

  6. Harvard–MIT Program in Health Sciences and Technology, Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA, USA

    Xining Gao

  7. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA

    Xining Gao, Micheal A. McLellan & Christopher S. Chen

  8. Department of Mechanical and Material Engineering, Florida International University, Miami, FL, USA

    Lihua Lou & Arvind Agarwal

  9. Department of Physics, Florida International University, Miami, FL, USA

    Jonathan Tabares & Jin He

  10. School of Engineering, Brown University, Providence, RI, USA

    Adriana C. Salazar Coariti & Kimani C. Toussaint Jr.

  11. Brown–Lifespan Center for Digital Health, Providence, RI, USA

    Kimani C. Toussaint Jr.

  12. Division of Materials Science and Engineering, Boston University, Boston, MA, USA

    Alice E. White

  13. Department of Physics, Boston University, Boston, MA, USA

    Alice E. White

Authors
  1. Jourdan K. Ewoldt
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  2. Samuel J. DePalma
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Contributions

J.K.E. and S.J.D. contributed equally, led and performed analysis, and wrote and reviewed the manuscript. B.M.B. and C.S.C. jointly supervised this work and wrote and reviewed the manuscript. M.E.J., M.Ç.K., Y.-M.L., P.M.H., X.G., L.L., M. McLellan, J.T., M. Ma and A.C.S.C. performed analysis and reviewed the manuscript. J.H., K.C.T., T.G.B., S.R., A.E.W., A.A. and E.L. supervised analysis and reviewed the manuscript.

Corresponding authors

Correspondence to Brendon M. Baker or Christopher S. Chen.

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Competing interests

C.S.C. is a founder and owns shares of Satellite Biosciences, a company that is developing cell-based therapies; and Ropirio Therapeutics, a company that is developing pharmaceuticals. All other authors declare no competing interests.

Peer review

Peer review information

Nature Methods thanks Nathan Palpant and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Primary Handling Editor: Madhura Mukhopadhyay, in collaboration with the Nature Methods team.

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Supplementary information

Supplementary Table 1

Top 15 reported hiPSC lines and their sex and ancestry. Information was obtained from publications, references or https://www.cellosaurus.org.

Supplementary Data 1

Complete dataset compiled from analysis of 300 studies using hiPSC-CM models for their selection of hiPSC lines, hiPSC-CM differentiation protocols, types of in vitro models, maturation techniques and metrics used to assess CM functionality and maturity. The dataset has been deposited at Dryad and will continue to be updated as new manuscripts are published in this area of research (https://www.doi.org/10.5061/dryad.ksn02v7bh).

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Ewoldt, J.K., DePalma, S.J., Jewett, M.E. et al. Induced pluripotent stem cell-derived cardiomyocyte in vitro models: benchmarking progress and ongoing challenges. Nat Methods 22, 24–40 (2025). https://doi.org/10.1038/s41592-024-02480-7

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