PhyMedExp, Université de Montpellier, INSERM, CNRS, 34295, Montpellier, France.
PhyMedExp, Université de Montpellier, INSERM, CNRS, CHU de Montpellier, 34295, Montpellier, France.
Biochem Biophys Res Commun. 2020 May 14;525(4):968-973. doi: 10.1016/j.bbrc.2020.03.020. Epub 2020 Mar 12.
Myogenic differentiation mechanisms are generally assessed using a murine cell line placed in low concentrations of an animal-derived serum. To more closely approximate in vivo pathophysiological conditions, recent studies have combined the use of human muscle cells with human serum. Nevertheless, the in vitro studies of the effects of a human microenvironment on the differentiation process of human myoblasts require the identification of the culture conditions that would provide an optimal and reproducible differentiation process of human muscle cells. We assessed the differentiation variability resulting from the use of human myoblasts and serums from healthy subjects by measuring the myotube diameter, fusion index and surface covered by myotubes. We showed the preserved cell-dependent variability of the differentiation response of myoblasts cultured in human serums compared to FBS. We found that using a pool of serums reduced the serum-dependent variability of the myogenic response compared to individual serums. We validated our methodology by showing the atrophying effect of pooled serums from COPD patients on healthy human myotubes. By replacing animal-derived tissues with human myoblasts and serums, and by validating the sensitivity of cultured human muscle cells to a pathological microenvironment, this human cell culture model offers a valuable tool for studying the role of the microenvironment in chronic disease.
成肌分化机制通常使用置于低浓度动物源性血清中的鼠细胞系来评估。为了更接近体内病理生理条件，最近的研究将人类肌肉细胞与人类血清结合使用。然而，在体外研究人类微环境对人类成肌细胞分化过程的影响需要确定提供人类肌肉细胞最佳和可重复分化过程的培养条件。我们通过测量肌管直径、融合指数和肌管覆盖的表面积，评估了使用健康供体的人类成肌细胞和血清导致的分化变异性。与 FBS 相比，我们表明在人血清中培养的成肌细胞的分化反应保持了细胞依赖性变异性。我们发现，与使用单个血清相比，使用血清池可降低肌生成反应的血清依赖性变异性。我们通过显示来自 COPD 患者的血清池对健康人肌管的萎缩作用验证了我们的方法学。通过用人成肌细胞和血清替代动物源性组织，并验证培养的人类肌肉细胞对病理微环境的敏感性，这种人类细胞培养模型为研究微环境在慢性疾病中的作用提供了有价值的工具。
