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Updated: Oct 11, 2025
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Fabrication of 3D Cardiac Microtissue Arrays using Human iPSC-Derived Cardiomyocytes, Cardiac Fibroblasts, and Endothelial Cells
Published on: March 14, 2021
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High-throughput 3D Spheroid Formation and Effective Cardiomyocyte Differentiation from Human iPS Cells Using the
Tatsuaki Miwa1, Alimjan Idiris2, Hiromichi Kumagai3
1Consumer Production Division, AGC Techno Glass Co., Ltd., Haibara-gun, Shizuoka, Japan.
Bio-Protocol
|December 3, 2021
Summary
This study introduces a novel method for generating uniform human induced pluripotent stem cell spheroids using EZSPHERE technology. This technique enhances cardiac differentiation, improving the purity and maturity of derived cardiomyocytes for regenerative medicine applications.
Area of Science:
- Stem Cell Biology
- Cardiovascular Research
- Biotechnology
Background:
- Human induced pluripotent stem cells (hiPSCs) are crucial for regenerative medicine and drug testing.
- Efficient generation of high-quality differentiated cells, like cardiomyocytes (hiPSC-CMs), is essential.
- Previous methods utilized microfabric vessels for hiPSC spheroid formation and differentiation, with Wnt signal control.
Purpose of the Study:
- To develop a simple, efficient protocol for large-scale, uniform hiPSC spheroid generation.
- To improve the cardiac differentiation process of hiPSCs using a spheroid re-aggregation method.
- To enhance the purity and maturity of hiPSC-derived cardiomyocytes.
Main Methods:
- Utilizing EZSPHERE microfabric vessels for high-throughput 3D spheroid formation from hiPSCs.
- Implementing a spheroid re-aggregation step during cardiac differentiation.
- Collecting, dissociating, and re-seeding single cells into EZSPHERE to form uniform hiPSC-CM spheroids.
Main Results:
- Successful generation of a large number of uniformly sized hiPSC spheroids.
- Demonstrated improvement in cardiac development through non-canonical Wnt signal promotion.
- Achieved enhanced purity and maturity of the resulting hiPSC-CMs.
Conclusions:
- The described re-aggregation protocol using EZSPHERE offers a simple and efficient method for cardiac differentiation.
- This approach significantly improves the quality of hiPSC-derived cardiomyocytes.
- The enhanced hiPSC-CMs hold promise for regenerative medicine and drug safety assessments.
