Micropost arrays for measuring stem cell-derived cardiomyocyte contractility

被引:77
作者
Beussman, Kevin M. [1 ]
Rodriguez, Marita L. [1 ]
Leonard, Andrea [1 ]
Taparia, Nikita [1 ]
Thompson, Curtis R. [1 ]
Sniadecki, Nathan J. [1 ,2 ]
机构
[1] Univ Washington, Dept Mech Engn, Seattle, WA 98195 USA
[2] Univ Washington, Dept Bioengn, Seattle, WA 98195 USA
基金
美国国家科学基金会;
关键词
Microposts; Induced pluripotent stem cells; Cardiomyocytes; Soft lithography; Cell mechanics; SINGLE CARDIOMYOCYTES; PROTEIN ADSORPTION; HEART FUNCTION; FORCE; MATURATION; ATTACHMENT; MANIPULATE; MECHANICS; IMPROVES; ADHESION;
D O I
10.1016/j.ymeth.2015.09.005
中图分类号
Q5 [生物化学];
学科分类号
070307 [化学生物学];
摘要
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. (C) 2015 Elsevier Inc. All rights reserved.
引用
收藏
页码:43 / 50
页数:8
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