Measuring the Contractile Forces of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes With Arrays of Microposts
Measuring the Contractile Forces of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes With Arrays of Microposts
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DOI：
10.1115/1.4027145
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发表时间：
2014-05-01
影响因子：
1.7
通讯作者：
Sniadecki, Nathan J.
中科院分区：
文献类型：
--
作者：
Rodriguez, Marita L.;Graham, Brandon T.;Sniadecki, Nathan J.
人类干细胞衍生的心肌细胞有望用于心脏修复、疾病建模、药物筛选和发育生物学研究。所有这些应用都可以通过在单细胞水平上评估心肌细胞的收缩性来改进。我们已经开发了一个体外平台，用于评估干细胞来源的心肌细胞的收缩性能，该平台与其他常见终点（如显微镜和分子生物学）兼容。将人诱导多能干细胞衍生的心肌细胞（hiPSC-CMs）植入弹性体微柱阵列，以表征这些细胞产生的收缩力、速度和能量。我们通过使用光学显微镜跟踪单个hiPSC-CM下微柱的偏转来评估收缩功能。对这些细胞进行免疫荧光染色，以评估其在微柱上的扩散面积、成核和肌肉结构。将hiPSC-CMs接种到包被纤维连接蛋白、层粘连蛋白和IV型胶原的微柱上，我们发现与包被纤维连接蛋白或IV型胶原的微柱相比，包被在层粘连蛋白涂层上的hiPSC-CMs表现出更高的附着、扩散面积和收缩速度。在优化条件下，hiPSC-CMs扩散面积约为420 mu m(2)，收缩力约为15 nN/ cells，收缩和松弛速率分别为1.74 mu m/s和1.46 mu m/s，峰值收缩功率为29 fW。因此，弹性体微柱阵列可以用于研究hiPSC-CMs的收缩强度和动力学。该系统将促进hiPSC-CM成熟、疾病建模、药物筛选以及人类心脏收缩的基础研究。
Human stem cell-derived cardiomyocytes hold promise for heart repair, disease modeling, drug screening, and for studies of developmental biology. All of these applications can be improved by assessing the contractility of cardiomyocytes at the single cell level. We have developed an in vitro platform for assessing the contractile performance of stem cell-derived cardiomyocytes that is compatible with other common endpoints such as microscopy and molecular biology. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were seeded onto elastomeric micropost arrays in order to characterize the contractile force, velocity, and power produced by these cells. We assessed contractile function by tracking the deflection of microposts beneath an individual hiPSC-CM with optical microscopy. Immunofluorescent staining of these cells was employed to assess their spread area, nucleation, and sarcomeric structure on the microposts. Following seeding of hiPSC-CMs onto microposts coated with fibronectin, laminin, and collagen IV, we found that hiPSC-CMs on laminin coatings demonstrated higher attachment, spread area, and contractile velocity than those seeded on fibronectin or collagen IV coatings. Under optimized conditions, hiPSC-CMs spread to an area of approximately 420 mu m(2), generated systolic forces of approximately 15 nN/cell, showed contraction and relaxation rates of 1.74 mu m/s and 1.46 mu m/s, respectively, and had a peak contraction power of 29 fW. Thus, elastomeric micropost arrays can be used to study the contractile strength and kinetics of hiPSC-CMs. This system should facilitate studies of hiPSC-CM maturation, disease modeling, and drug screens as well as fundamental studies of human cardiac contraction.