iBB - Institute for Bioengineering and Biosciences and Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisbon, Portugal; The Discoveries Centre for Regenerative and Precision Medicine, Lisbon Campus, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisbon, Portugal.
iBB - Institute for Bioengineering and Biosciences and Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisbon, Portugal; Centro de Química-Física Molecular and Institute of Nanoscience and Nanotechnology, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisbon, Portugal.
Biochem Biophys Res Commun. 2018 May 15;499(3):611-617. doi: 10.1016/j.bbrc.2018.03.198. Epub 2018 Mar 30.
Human induced Pluripotent Stem Cell-derived cardiomyocytes (hiPSC-CMs) have an enormous potential for the development of drug screening and modeling cardiac disease platforms. However, early hiPSC-CMs usually exhibit low structural development, precluding the applicability of these cells. Here, we follow during 120 days the progressive structural maturation of hiPSC-CM microtissues obtained using the Wnt signaling modulation protocol. For this purpose, we designed a user friendly custom-written program to quantify cardiac fiber alignment and sarcomere length. Cardiomyocyte shape, cardiac fiber density and multinucleation were also analyzed. Derived cardiomyocytes showed significant progression in cardiomyocyte fiber density and sarcomere length during the long-term culture, with a peak at day 90 of 40% multinucleated cells. In addition, cardiomyocyte microtissues remained functional with progressive maturation leading to a decrease in the percentage of cTnT positive cells from 59% to 22% at day 120, a value similar to the content present in tissues of the adult left ventricle. These data and the framework that we provide to quantify cardiomyocyte structural features can be important to set new metrics to develop applications for drug screening and disease modeling.
人诱导多能干细胞衍生的心肌细胞（hiPSC-CMs）在药物筛选和心脏疾病模型平台的开发方面具有巨大的潜力。然而，早期的 hiPSC-CMs 通常表现出低结构发育，限制了这些细胞的适用性。在这里，我们在 120 天的时间里跟踪了使用 Wnt 信号调节方案获得的 hiPSC-CM 微组织的渐进结构成熟过程。为此，我们设计了一个用户友好的自定义程序来量化心肌纤维取向和肌节长度。还分析了心肌细胞的形状、心肌纤维密度和多核化。在长期培养过程中，衍生的心肌细胞的心肌纤维密度和肌节长度显著增加，在第 90 天达到 40%的多核细胞峰值。此外，随着成熟的进行，心肌细胞微组织保持功能，导致第 120 天 cTnT 阳性细胞的百分比从 59%下降到 22%，这一值与成年左心室组织中的含量相似。这些数据和我们提供的量化心肌细胞结构特征的框架对于制定新的指标来开发药物筛选和疾病建模应用程序可能非常重要。
