Matrix-Patterned Micromarkers for Tracking Local Contractile Movements of Myotubes Cultured on Thin-Film Scaffold

S. Hashimoto
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引用次数: 1

Abstract

To measure the distribution of local contractile movements in the layer of the myotubes, the movement of the matrix pattern of micro-markers placed on the back surface of the myotube-attached thin film has been tracked in vitro. A matrix pattern of micromarkers (diameter: 4 μm, height: 2 μm, interval: 30 μm) was fabricated on the back of a polydimethylsiloxane thin film (thickness: 6 μm) using photolithography. Mouse myoblasts were seeded on the front of the thin film and cultured for 10 days to differentiate into myotubes. Periodic electrical pulses (amplitude 2 V, pulse period 0.5 s, pulse width 1 ms) were applied between the electrodes of titanium wires immersed in the medium. Movements of both myotubes and markers were observed simultaneously with an optical microscope. Experimental results indicate that the motion distribution of each marker is related to the local periodic motion of the myotube layer. Thin films with matrix-patterned micromarkers fabricated on the back side allow in vitro analysis of the distribution of local repetitive contractile movements in the myotube layer.
基质模式微标记跟踪薄膜支架上肌管局部收缩运动
为了测量肌管层局部收缩运动的分布,我们在体外跟踪了放置在肌管附着薄膜背面的微标记物基质图案的运动。采用光刻技术在厚度为6 μm的聚二甲基硅氧烷薄膜背面制备了直径为4 μm、高度为2 μm、间距为30 μm的微标记基质图案。将小鼠成肌细胞播种于薄膜前部,培养10天分化成肌管。在浸泡在介质中的钛丝电极之间施加周期电脉冲(振幅2 V,脉冲周期0.5 s,脉冲宽度1 ms)。用光学显微镜同时观察肌管和标记物的运动。实验结果表明,各标记物的运动分布与肌管层的局部周期性运动有关。在背面制造的带有基质图案微标记的薄膜允许在体外分析肌管层局部重复收缩运动的分布。
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