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Updated: Oct 11, 2025
Pluripotent Stem Cell Derived Cardiac Cells for Myocardial Repair
Published on: February 3, 2017
Replating Protocol for Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
Arzuhan Koc1,2, Esra Cagavi3,4,5
1Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, Turkey.
Insights
Optimized protocols for replating human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) ensure high viability. These methods effectively generate single-cell or cluster hiPSC-CM cultures for cardiac research.
Area of Science:
- Cardiovascular Biology
- Stem Cell Biology
- Biotechnology
Background:
- Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) offer a renewable source for cardiac research.
- Challenges exist in obtaining hiPSC-CM cultures in desired formats (single-cell, monolayer, 3D clusters) for downstream applications.
Purpose of the Study:
- To develop and optimize replating strategies for hiPSC-CMs.
- To evaluate various dissociation reagents and matrix coatings for effective reseeding.
Main Methods:
- Dissociation of hiPSC-CMs using collagenase A&B, Collagenase II, TrypLE, and EDTA.
- Reseding of dissociated hiPSC-CMs on matrix materials including fibronectin, laminin, iMatrix, Matrigel, and Geltrex.
Main Results:
- Defined optimized protocols for hiPSC-CM dissociation and reseeding.
- Achieved high viability of reseeding hiPSC-CMs.
- Successfully generated single-cell or cluster-containing hiPSC-CM cultures.
Conclusions:
- Optimized replating strategies are critical for generating functional hiPSC-CM cultures.
- The described methods provide effective means to obtain hiPSC-CMs in desired formats for diverse research applications.
- These protocols enhance the utility of hiPSC-CMs in basic and translational cardiac research.
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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