Departments of Mechanical Engineering and of Bioengineering, Stanford University, School of Engineering and School of Medicine, Stanford, CA, USA.
Baxter Laboratory for Stem Cell Biology, Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA, USA.
Nat Commun. 2024 Jun 26;15(1):5427. doi: 10.1038/s41467-024-49755-3.
Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) are powerful in vitro models to study the mechanisms underlying cardiomyopathies and cardiotoxicity. Quantification of the contractile function in single hiPSC-CMs at high-throughput and over time is essential to disentangle how cellular mechanisms affect heart function. Here, we present CONTRAX, an open-access, versatile, and streamlined pipeline for quantitative tracking of the contractile dynamics of single hiPSC-CMs over time. Three software modules enable: parameter-based identification of single hiPSC-CMs; automated video acquisition of >200 cells/hour; and contractility measurements via traction force microscopy. We analyze >4,500 hiPSC-CMs over time in the same cells under orthogonal conditions of culture media and substrate stiffnesses; +/- drug treatment; +/- cardiac mutations. Using undirected clustering, we reveal converging maturation patterns, quantifiable drug response to Mavacamten and significant deficiencies in hiPSC-CMs with disease mutations. CONTRAX empowers researchers with a potent quantitative approach to develop cardiac therapies.
人诱导多能干细胞（hiPSC-CMs）衍生的心肌细胞是研究心肌病和心脏毒性的潜在机制的强大体外模型。高通量和长时间内对单个 hiPSC-CMs 的收缩功能进行定量分析对于阐明细胞机制如何影响心脏功能至关重要。在这里，我们提出了 CONTRAX，这是一个用于定量跟踪单个 hiPSC-CMs 随时间收缩动态的开放访问、多功能且简化的流水线。三个软件模块实现了：基于参数的单个 hiPSC-CMs 的识别；>200 个细胞/小时的自动视频采集；以及通过牵引力显微镜进行收缩力测量。我们在相同细胞中，在正交的培养基和基质硬度条件下；药物处理的 +/-；心脏突变的 +/- 下，随时间分析了>4500 个 hiPSC-CMs。使用无向聚类，我们揭示了趋同的成熟模式、对 Mavacamten 的可量化药物反应以及疾病突变的 hiPSC-CMs 明显缺陷。CONTRAX 为研究人员提供了一种强大的定量方法来开发心脏治疗方法。
