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Replating Protocol for Human Induced Pluripotent Stem Cell–Derived Cardiomyocytes

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Embryonic Stem Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2520))

Abstract

Human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CM) create an unlimited cell source for basic and translational cardiac research. Obtaining hiPSC-CM culture as a single-cell, monolayer or three-dimensional clusters for downstream applications can be challenging. Thus, it is critical to develop replating strategies for hiPSC-CMs by evaluating different enzymatic or nonenzymatic reagents for dissociation and seeding on different coating materials. To reseed hiPSC-CMs with high viability and at structures desirable for the downstream applications, here we defined optimized protocols to dissociate hiPSC-CMs by using collagenase A&B, Collagenase II, TrypLE, and EDTA and reseeding on various matrix materials including fibronectin, laminin, imatrix, Matrigel, and Geltrex. By the replating methods described here, a single cell or cluster-containing hiPSC-CM cultures can be generated effectively.

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Abbreviations

ECM:

Extracellular matrix

EDTA:

Ethylenediaminetetraacetic acid

hiPSC-CM:

Human induced pluripotent stem cell-derived cardiomyocytes

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Acknowledgments

This work was supported by the Scientific and Technological Research Council of Turkey (TÜBİTAK) under grant number 213S192.

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Authors and Affiliations

  1. Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, Turkey

    Arzuhan Koc & Esra Cagavi

  2. Medical Microbiology Graduate Program, Health Sciences Institute, Istanbul Medipol University, Istanbul, Turkey

    Arzuhan Koc

  3. Department of Medical Biology, School of Medicine, Istanbul Medipol University, Istanbul, Turkey

    Esra Cagavi

  4. Medical Biology and Genetics Graduate Program, Health Sciences Institute, Istanbul Medipol University, Istanbul, Turkey

    Esra Cagavi

Authors
  1. Arzuhan Koc
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  2. Esra Cagavi
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Corresponding author

Correspondence to Esra Cagavi .

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Editors and Affiliations

  1. Ottawa, ON, Canada

    Kursad Turksen

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Koc, A., Cagavi, E. (2021). Replating Protocol for Human Induced Pluripotent Stem Cell–Derived Cardiomyocytes. In: Turksen, K. (eds) Embryonic Stem Cell Protocols . Methods in Molecular Biology, vol 2520. Humana, New York, NY. https://doi.org/10.1007/7651_2021_450

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  • DOI: https://doi.org/10.1007/7651_2021_450

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2436-4

  • Online ISBN: 978-1-0716-2437-1

  • eBook Packages: Springer Protocols

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