Wan Zhongjun, Wen Shanshan, Zheng Ran, Li Li, Jiang Wei, Zhang Donghui
State Key Laboratory of Biocatalysis and Enzyme Engineering, Stem Cells and Tissue Engineering Manufacture Center, School of Life Sciences, Hubei University, Wuhan 430062, China.
Department of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, China.
STAR Protoc. 2025 Mar 21;6(1):103576. doi: 10.1016/j.xpro.2024.103576. Epub 2025 Jan 31.
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. and Li et al..
扩展多能干细胞（EPSCs）具有高分化能力，有可能作为生成心肌细胞的优质种子资源。在此，我们展示了一种用于生成无饲养层EPSCs（ffEPSCs）、心肌细胞和工程心脏组织（EHTs）的方案。我们描述了将人类胚胎干细胞或诱导多能干细胞（ESCs/iPSCs）转化为ffEPSCs的步骤，随后是它们的长期维持、冷冻保存、种子保存以及分化为心肌细胞。然后我们详细介绍构建和培养三维EHTs的程序，随后进行其收缩力测量和光学映射。有关该方案使用和执行的完整详细信息，请参考郑等人和李等人的研究。
