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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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
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2436-4
Online ISBN: 978-1-0716-2437-1
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