Protocol for differentiating cardiomyocytes and generating engineered heart tissues from human feeder-free extended pluripotent stem cells
Summary: Extended pluripotent stem cells (EPSCs) possess a high differentiation capacity, potentially as a superior seed resource for generating cardiomyocytes. Here, we present a protocol for generating feeder-free EPSCs (ffEPSCs), cardiomyocytes, and engineered heart tissues (EHTs). We describe st...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672400741X |
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| author | Zhongjun Wan Shanshan Wen Ran Zheng Li Li Wei Jiang Donghui Zhang |
| author_facet | Zhongjun Wan Shanshan Wen Ran Zheng Li Li Wei Jiang Donghui Zhang |
| author_sort | Zhongjun Wan |
| collection | DOAJ |
| description | Summary: Extended pluripotent stem cells (EPSCs) possess a high differentiation capacity, potentially as a superior seed resource for generating cardiomyocytes. Here, we present a protocol for generating feeder-free EPSCs (ffEPSCs), cardiomyocytes, and engineered heart tissues (EHTs). We describe steps for converting human embryonic stem cells or induced pluripotent stem cells (ESCs/iPSCs) into ffEPSCs, followed by their long-term maintenance, cryopreservation, seed preservation, and differentiation into cardiomyocytes. We then detail procedures for constructing and culturing three-dimensional EHTs followed by their contraction force measurement and optical mapping.For complete details on the use and execution of the protocol, please refer to Zheng et al.1 and Li et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-644d9463490d4318ab2696335dca6c0c |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-644d9463490d4318ab2696335dca6c0c2025-02-02T05:29:20ZengElsevierSTAR Protocols2666-16672025-03-0161103576Protocol for differentiating cardiomyocytes and generating engineered heart tissues from human feeder-free extended pluripotent stem cellsZhongjun Wan0Shanshan Wen1Ran Zheng2Li Li3Wei Jiang4Donghui Zhang5State Key Laboratory of Biocatalysis and Enzyme Engineering, Stem Cells and Tissue Engineering Manufacture Center, School of Life Sciences, Hubei University, Wuhan 430062, ChinaDepartment of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, ChinaDepartment of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, ChinaState Key Laboratory of Biocatalysis and Enzyme Engineering, Stem Cells and Tissue Engineering Manufacture Center, School of Life Sciences, Hubei University, Wuhan 430062, ChinaDepartment of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, China; Corresponding authorState Key Laboratory of Biocatalysis and Enzyme Engineering, Stem Cells and Tissue Engineering Manufacture Center, School of Life Sciences, Hubei University, Wuhan 430062, China; Corresponding authorSummary: Extended pluripotent stem cells (EPSCs) possess a high differentiation capacity, potentially as a superior seed resource for generating cardiomyocytes. Here, we present a protocol for generating feeder-free EPSCs (ffEPSCs), cardiomyocytes, and engineered heart tissues (EHTs). We describe steps for converting human embryonic stem cells or induced pluripotent stem cells (ESCs/iPSCs) into ffEPSCs, followed by their long-term maintenance, cryopreservation, seed preservation, and differentiation into cardiomyocytes. We then detail procedures for constructing and culturing three-dimensional EHTs followed by their contraction force measurement and optical mapping.For complete details on the use and execution of the protocol, please refer to Zheng et al.1 and Li et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S266616672400741XCell cultureStem CellsCell DifferentiationOrganoidsTissue Engineering |
| spellingShingle | Zhongjun Wan Shanshan Wen Ran Zheng Li Li Wei Jiang Donghui Zhang Protocol for differentiating cardiomyocytes and generating engineered heart tissues from human feeder-free extended pluripotent stem cells STAR Protocols Cell culture Stem Cells Cell Differentiation Organoids Tissue Engineering |
| title | Protocol for differentiating cardiomyocytes and generating engineered heart tissues from human feeder-free extended pluripotent stem cells |
| title_full | Protocol for differentiating cardiomyocytes and generating engineered heart tissues from human feeder-free extended pluripotent stem cells |
| title_fullStr | Protocol for differentiating cardiomyocytes and generating engineered heart tissues from human feeder-free extended pluripotent stem cells |
| title_full_unstemmed | Protocol for differentiating cardiomyocytes and generating engineered heart tissues from human feeder-free extended pluripotent stem cells |
| title_short | Protocol for differentiating cardiomyocytes and generating engineered heart tissues from human feeder-free extended pluripotent stem cells |
| title_sort | protocol for differentiating cardiomyocytes and generating engineered heart tissues from human feeder free extended pluripotent stem cells |
| topic | Cell culture Stem Cells Cell Differentiation Organoids Tissue Engineering |
| url | http://www.sciencedirect.com/science/article/pii/S266616672400741X |
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