BACKGROUND Formation of muscular pseudo-tissue depends on muscle precursor cells, the extracellular matrix (ECM)-mimicking structure and factors stimulating cell differentiation. These three things cooperate and can create a tissue-like structure, however, their interrelationships are relatively unknown. The objective was to study the interaction between surface properties, culture medium composition and heterogeneous cell culture. We would like to demonstrate that changing the surface properties by coating with graphene oxide nanofilm (nGO) can affect cell behaviour and especially their need for the key amino acid L-glutamine (L-Glu). RESULTS Chicken embryo muscle cells and their precursors, cultured in vitro, were used as the experimental model. The mesenchymal stem cell, collected from the hind limb of the chicken embryo at day 8 were divided into 4 groups; the control group and groups treated with nGO, L-Glu and nGO supplied with L-Glu (nGOxL-Glu). The roughness of the surface of the plastic plate covered with nGO was much lower than a standard plate. The test of nGO biocompatibility demonstrated that the cells were willing to settle on the nGO without any toxic effects. Moreover, nGO by increasing hydrophilicity and reducing roughness and presumably through chemical bonds available on the GO surface stimulated the colonisation of primary stromal cells that promote embryonic satellite cells. The viability significantly increased in cells cultured on nGOxL-Glu. Observations of cell morphology showed that the most mature state of myogenesis was characteristic for the group nGOxL-Glu. This result was confirmed by increasing the expression of MYF5 genes at mRNA and protein levels. nGO also increased the expression of MYF5 and also very strongly the expression of PAX7 at mRNA and protein levels. However, when analysing the expression of PAX7, a positive link was observed between the nGO surface and the addition of L-Glu. CONCLUSIONS The use of nGO and L-Glu supplement may improve myogenesis and also the myogenic potential of myocytes and their precursors by promoting the formation of satellite cells. Studies have, for the first time, demonstrated positive cooperation between surface properties nGO and L-Glu supplementation to the culture medium regarding the myogenic potential of cells involved in muscle formation.

中文翻译：

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氧化石墨烯纳米膜和L-谷氨酰胺的添加可以促进胚胎肌肉细胞的发育。

背景技术肌肉假组织的形成取决于肌肉前体细胞，模拟细胞外基质（ECM）的结构和刺激细胞分化的因素。这三件事相互配合，可以创建一个组织状的结构，但是它们之间的相互关系是相对未知的。目的是研究表面性质，培养基组成和异质细胞培养之间的相互作用。我们想证明通过涂以氧化石墨烯纳米膜（nGO）来改变表面性能会影响细胞行为，尤其是对关键氨基酸L-谷氨酰胺（L-Glu）的需求。结果体外培养的鸡胚肌细胞及其前体细胞被用作实验模型。间充质干细胞 在第8天从鸡胚的后肢收集的被分成4组。对照组和接受nGO，L-Glu和nGO供给L-Glu（nGOxL-Glu）治疗的组。用nGO覆盖的塑料板的表面粗糙度要比标准板低得多。对nGO生物相容性的测试表明，细胞愿意在无任何毒性作用的情况下沉淀在nGO上。而且，nGO通过增加亲水性和降低粗糙度，并大概通过GO表面上可用的化学键，刺激了促进胚胎卫星细胞的原代基质细胞的定殖。在nGOxL-Glu上培养的细胞中，活力显着提高。细胞形态的观察表明，最成熟的肌发生状态是nGOxL-Glu组的特征。通过在mRNA和蛋白质水平上增加MYF5基因的表达，证实了这一结果。nGO在mRNA和蛋白水平上也增加了MYF5的表达，也非常强烈地增强了PAX7的表达。但是，在分析PAX7的表达时，在nGO表面与添加L-Glu之间观察到正向联系。结论使用nGO和L-Glu补充剂可通过促进卫星细胞的形成来改善肌细胞及其前体的肌发生能力，并改善其成肌潜能。研究首次表明，关于参与肌肉形成的细胞的成肌潜能，nGO表面特性和向培养基中添加L-Glu之间存在积极的合作关系。nGO在mRNA和蛋白水平上也增加了MYF5的表达，也非常强烈地增强了PAX7的表达。但是，在分析PAX7的表达时，在nGO表面与添加L-Glu之间观察到正向联系。结论使用nGO和L-Glu补充剂可以通过促进卫星细胞的形成来改善肌细胞及其前体的肌发生潜能。研究首次表明，关于参与肌肉形成的细胞的成肌潜能，nGO表面特性和向培养基中添加L-Glu之间存在积极的合作关系。nGO在mRNA和蛋白水平上也增加了MYF5的表达，也非常强烈地增强了PAX7的表达。但是，在分析PAX7的表达时，在nGO表面与添加L-Glu之间观察到正向联系。结论使用nGO和L-Glu补充剂可以通过促进卫星细胞的形成来改善肌细胞及其前体的肌发生潜能。研究首次表明，关于参与肌肉形成的细胞的成肌潜力，nGO表面特性和向培养基中添加L-Glu之间存在积极的合作关系。结论使用nGO和L-Glu补充剂可通过促进卫星细胞的形成来改善肌细胞及其前体的肌发生能力，并改善其成肌潜能。研究首次表明，关于参与肌肉形成的细胞的成肌潜能，nGO表面特性和向培养基中添加L-Glu之间存在积极的合作关系。结论使用nGO和L-Glu补充剂可以通过促进卫星细胞的形成来改善肌细胞及其前体的肌发生潜能。研究首次表明，关于参与肌肉形成的细胞的成肌潜力，nGO表面特性和向培养基中添加L-Glu之间存在积极的合作关系。