Micropost arrays for measuring stem cell-derived cardiomyocyte contractility.
Micropost arrays for measuring stem cell-derived cardiomyocyte contractility.
复制标题
DOI：
10.1016/j.ymeth.2015.09.005
复制
发表时间：
2016-02-01
期刊：
影响因子：
--
通讯作者：
Sniadecki NJ
中科院分区：
文献类型：
--
作者：
Beussman KM;Rodriguez ML;Leonard A;Taparia N;Thompson CR;Sniadecki NJ
干细胞衍生的心肌细胞具有用于研究心脏疾病和成熟、筛选药物治疗和恢复心脏功能的潜力。在这里，我们讨论了使用微柱阵列来测量干细胞衍生的心肌细胞产生的收缩力所涉及的程序。心脏泵血需要心肌细胞的收缩力，因此测量心肌细胞的收缩力是评估其功能的直接方法。利用微加工和软光刻技术从共同的母版创建相同的柔性硅树脂微柱阵列。微柱阵列用细胞外基质蛋白功能化以允许心肌细胞粘附到微柱的尖端。实时成像用于捕获由心肌细胞收缩引起的微柱偏转的视频。图像分析代码提供了一种精确的手段来量化这些偏转。通过将微柱建模为悬臂梁来计算由跳动的心肌细胞产生的收缩力。我们已经使用该测定来评估用于改善干细胞衍生的心肌细胞的成熟和收缩功能的技术。
Stem cell-derived cardiomyocytes have the potential to be used to study heart disease and maturation, screen drug treatments, and restore heart function. Here, we discuss the procedures involved in using micropost arrays to measure the contractile forces generated by stem cell-derived cardiomyocytes. Cardiomyocyte contractility is needed for the heart to pump blood, so measuring the contractile forces of cardiomyocytes is a straightforward way to assess their function. Microfabrication and soft lithography techniques are utilized to create identical arrays of flexible, silicone microposts from a common master. Micropost arrays are functionalized with extracellular matrix protein to allow cardiomyocytes to adhere to the tips of the microposts. Live imaging is used to capture videos of the deflection of microposts caused by the contraction of the cardiomyocytes. Image analysis code provides an accurate means to quantify these deflections. The contractile forces produced by a beating cardiomyocyte are calculated by modeling the microposts as cantilever beams. We have used this assay to assess techniques for improving the maturation and contractile function of stem cell-derived cardiomyocytes.