Abstract：
The adept and systematic differentiation of embryonic stem cells (ESCs) and human-induced pluripotent stem cells (hiPSCs) to diverse lineage-prone cell types involves crucial step-by-step process that mimics the vital strategic commitment phase that is usually observed during the process of embryo development. The development of precise tissue-specific cell types from these stem cells indeed plays an important role in the advancement of imminent stem cell-based therapeutic strategies. Therefore, the usage of hiPSC-derived cell types for subsequent cardiovascular disease modeling, drug screening, and therapeutic drug development undeniably entails an in-depth understanding of each and every step to proficiently stimulate these stem cells into desired cardiomyogenic lineage. Thus, to accomplish this definitive and decisive fate, it is essential to efficiently induce the mesoderm or pre-cardiac mesoderm, succeeded by the division of cells into cardiovascular and ultimately ensuing with the cardiomyogenic lineage outcome. This usually commences from the earliest phases of pluripotent cell induction. In this chapter, we discuss our robust and reproducible step-wise protocol that will describe the subtype controlled, precise lineage targeted standardization of activin/nodal, and BMP signaling molecules/cytokines, for the efficient differentiation of ventricular cardiomyocytes from hiPSCs via the embryoid body method. In addition, we also describe techniques to dissociate hiPSCs, hiPSC-derived early cardiomyocytes for mesoderm and pre-cardiac mesoderm assessment, and hiPSC-derived cardiomyocytes for early and mature markers assessment.
摘要：
胚胎干细胞（ESC）和人类诱导的多能干细胞（hiPSC）向多种谱系易发细胞类型的熟练和系统分化涉及关键的逐步过程，该过程模拟了通常在胚胎发育过程中观察到的重要战略承诺阶段。从这些干细胞开发精确的组织特异性细胞类型确实在即将到来的基于干细胞的治疗策略的发展中起着重要作用。因此,在随后的心血管疾病建模中使用hiPSC衍生的细胞类型，药物筛选,不可否认，治疗药物的开发需要深入了解每一步，以熟练地刺激这些干细胞进入所需的心肌谱系。因此，为了完成这一决定性的命运，有效诱导中胚层或前心脏中胚层是必不可少的，成功的细胞分裂成心血管，并最终与心肌谱系的结果。这通常从多能细胞诱导的最早阶段开始。在这一章中,我们讨论了我们的健壮和可重复的分步方案，该方案将描述受控的亚型，激活素/节点的精确谱系目标标准化，和BMP信号分子/细胞因子，通过胚体方法从hiPSCs有效分化心室心肌细胞。此外,我们还描述了分离HiPSCs的技术，hiPSC衍生的早期心肌细胞用于中胚层和心脏中胚层前评估，和hiPSC衍生的心肌细胞用于早期和成熟标志物评估。
