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首页> 外文期刊>Cytoskeleton >Combined substrate micropatterning and FFT analysis reveals myotube size control and alignment by contact guidance
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Combined substrate micropatterning and FFT analysis reveals myotube size control and alignment by contact guidance

机译：组合的衬底微型仪器和FFT分析显示MyoTube尺寸控制和通过联系指导对准

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Abstract 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.
机译：摘要最重要的评价体外骨骼肌模型的标准包括分化的程度和组织模型中的对准程度。基板微图案被认为是一种有效的工具，因为它在体内重现如细胞微环境，并有助于理解肌生成的基本概念和机制。然而，需要进一步探索基于微型接触引导提示的基于卫星细胞对准和肌室形成的影响。在目前的工作中，我们证明了肌管尺寸控制的调节并通过基板微图案对准。为此目的，从大鼠后肢肌肉中分离的原发性肌细胞细胞（即，卫星细胞）的特征和培养为通过微池流动的等离子体工艺处理的微型理由的玻璃基板上的14天的时间。量化了几种肌肉分化的特征参数，包括熔融指数，成熟指数和肌管的平均宽度。通过脑识别进行评估表现出自发收缩的培养的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Myotubes对微图案基板上的对准程度。总的来说，结果表明，由于基材的微图案而产生的接触引导提示可以是用于控制肌发育过程中肌管的尺寸和程度的关键调节因子。

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