Sun Xuetao, Nunes Sara S
Toronto General Research Institute, University Health Network.
Toronto General Research Institute, University Health Network; Institute of Biomaterials and Biomedical Engineering, University of Toronto; Heart & Stroke/Richard Lewar Centre of Excellence, University of Toronto; Laboratory of Medicine and Pathobiology, University of Toronto;
J Vis Exp. 2017 May 6(123):55373. doi: 10.3791/55373.
Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have been a promising cell source and have thus encouraged the investigation of their potential applications in cardiac research, including drug discovery, disease modeling, tissue engineering, and regenerative medicine. However, cells produced by existing protocols display a range of immaturity compared with native adult ventricular cardiomyocytes. Many efforts have been made to mature hPSC-CMs, with only moderate maturation attained thus far. Therefore, an engineered system, called biowire, has been devised by providing both physical and electrical cues to lead hPSC-CMs to a more mature state in vitro. The system uses a microfabricated platform to seed hPSC-CMs in collagen type I gel along a rigid template suture to assemble into aligned cardiac tissue (biowire), which is subjected to electrical field stimulation with a progressively increasing frequency. Compared to nonstimulated controls, stimulated biowired cardiomyocytes exhibit an enhanced degree of structural and electrophysiological maturation. Such changes are dependent upon the stimulation rate. This manuscript describes in detail the design and creation of biowires.
人多能干细胞衍生的心肌细胞（hPSC-CMs）一直是一种很有前景的细胞来源，因此激发了人们对其在心脏研究中潜在应用的探索，包括药物发现、疾病建模、组织工程和再生医学。然而，与天然成年心室心肌细胞相比，现有方案产生的细胞表现出一系列不成熟的特征。人们已经做出了许多努力来使hPSC-CMs成熟，到目前为止仅实现了适度的成熟。因此，一种名为生物线的工程系统被设计出来，通过提供物理和电信号线索，引导hPSC-CMs在体外达到更成熟的状态。该系统使用一个微加工平台，将hPSC-CMs沿着刚性模板缝线接种到I型胶原凝胶中，组装成排列整齐的心脏组织（生物线），并对其施加频率逐渐增加的电场刺激。与未受刺激的对照组相比，受刺激的生物线心肌细胞在结构和电生理成熟度上有增强。这些变化取决于刺激速率。本手稿详细描述了生物线的设计和创建。
