Most important evaluating criteria for in vitro skeletal muscle models include the extent of differentiation and the degree of alignment in the tissue model. Substrate micropatterning is considered as an effective tool as it recreates in vivo like cellular microenvironment and helps in understanding the fundamental concepts and mechanisms underlying myogenesis. However, the influence of micropatterning based contact guidance cues over satellite cell alignment and myotube formation needs to be explored and studied further. In the present work, we demonstrate the regulation of myotube size control and alignment through the substrate micropatterning. For this purpose, primary myoblast cells (i.e., satellite cells) isolated from rat hind limb muscle were characterized and cultured for a period of 14 days on micropatterned glass substrates processed by the microchannnel flowed plasma process. Several characteristic parameters of muscle differentiation, including the fusion index, maturation index, and average width of the myotubes were quantified. The functional behavior of cultured myotubes exhibiting spontaneous contractions was assessed through kymograph to determine the twitch frequency. In addition, we evaluated the degree of alignment of myotubes on micropatterned substrates through examining orientation order parameter and two‐dimensional fast Fourier transform analysis. Altogether, the outcomes reveal that the contact guidance cues arising due to micropatterning of the substrates could be a key regulator for controlling the size and degree of alignment of myotubes during the myogenesis process.

中文翻译：

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结合的基板微图案和FFT分析揭示了肌管尺寸控制和通过接触引导的对准。

体外骨骼肌模型最重要的评估标准包括组织模型中的分化程度和对齐程度。底物微图案化被认为是一种有效的工具，因为它可以像细胞微环境一样在体内重建，并有助于理解成肌的基本概念和机制。然而，基于微模式的接触指导线索对卫星细胞对准和肌管形成的影响需要进一步研究。在目前的工作中，我们演示了通过基质微图案对肌管大小控制和对齐的调节。为此，原代成肌细胞（即 对从大鼠后肢肌肉分离的卫星细胞进行表征，并在通过微通道流动等离子体工艺处理的微图案化玻璃基板上培养14天。量化了肌肉分化的几个特征参数，包括融合指数，成熟指数和肌管的平均宽度。通过运动记录仪评估显示自发收缩的培养肌管的功能行为，以确定抽搐频率。此外，我们通过检查取向顺序参数和二维快速傅里叶变换分析，评估了微管基质上肌管的排列程度。共，