Beussman Kevin M, Rodriguez Marita L, Leonard Andrea, Taparia Nikita, Thompson Curtis R, Sniadecki Nathan J
Department of Mechanical Engineering, University of Washington, Seattle, WA, USA.
Department of Mechanical Engineering, University of Washington, Seattle, WA, USA; Department of Bioengineering, University of Washington, Seattle, WA, USA.
Methods. 2016 Feb 1;94:43-50. doi: 10.1016/j.ymeth.2015.09.005. Epub 2015 Sep 3.
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.
干细胞衍生的心肌细胞有潜力用于研究心脏病与心肌成熟、筛选药物治疗方法以及恢复心脏功能。在此，我们讨论使用微柱阵列来测量干细胞衍生心肌细胞产生的收缩力所涉及的步骤。心脏泵血需要心肌细胞的收缩性，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，从一个通用模具制作出相同的柔性硅微柱阵列。微柱阵列用细胞外基质蛋白进行功能化处理，以使心肌细胞能够附着在微柱的尖端。通过实时成像来捕捉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准确量化这些偏转的方法。通过将微柱建模为悬臂梁来计算跳动的心肌细胞产生的收缩力。我们已使用该测定法来评估改善干细胞衍生心肌细胞成熟度和收缩功能的技术。
