Zhu Wei-Zhong, Van Biber Benjamin, Laflamme Michael A
Laflamme Lab, Pathology, Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, USA.
Methods Mol Biol. 2011;767:419-31. doi: 10.1007/978-1-61779-201-4_31.
The availability of human cardiomyocytes derived from embryonic stem cells (ESCs) has generated -considerable excitement, as these cells are an excellent model system for studying myocardial development and may have eventual application in cell-based cardiac repair. Cardiomyocytes derived from the related induced pluripotent stem cells (iPSCs) have similar properties, but also offer the prospects of patient-specific disease modeling and cell therapies. Unfortunately, the methods by which cardiomyocytes have been historically generated from pluripotent stem cells are unreliable and typically result in preparations of low cardiac purity (typically <1% cardiomyocytes). We detail here the methods for a recently reported directed cardiac differentiation protocol, which involves the serial application of two growth factors known to be involved in early embryonic heart development, activin A, and bone morphogenetic protein-4 (BMP-4). This protocol reliably yields preparations of 30-60% cardiomyocytes, which can then be further enriched to >90% cardiomyocytes using straightforward physical methods.
源自胚胎干细胞（ESC）的人类心肌细胞的可得性引发了极大的关注，因为这些细胞是研究心肌发育的优秀模型系统，并且最终可能应用于基于细胞的心脏修复。源自相关诱导多能干细胞（iPSC）的心肌细胞具有相似的特性，但也为患者特异性疾病建模和细胞疗法提供了前景。不幸的是，历史上从多能干细胞生成心肌细胞的方法不可靠，通常导致心脏纯度较低的制剂（通常<1%心肌细胞）。我们在此详细介绍一种最近报道的定向心脏分化方案的方法，该方案涉及依次应用两种已知参与早期胚胎心脏发育的生长因子，激活素A和骨形态发生蛋白-4（BMP-4）。该方案可靠地产生30-60%心肌细胞的制剂，然后可以使用简单的物理方法进一步富集到>90%的心肌细胞。
