Univ Grenoble Alpes, CNRS, INSERM, IAB, Allée des Alpes, 38000, Grenoble, France.
Univ Lyon, CNRS, Université Claude Bernard Lyon 1, ENS de Lyon, Institut des Sciences Analytiques, UMR 5280, 5 rue de la Doua, 69100, Villeurbanne, France.
Sci Rep. 2020 Jan 31;10(1):1622. doi: 10.1038/s41598-020-58377-w.
Stem cells, poised to revolutionize current medicine, stand as major workhorses for monitoring changes in cell fate. Characterizing metabolic phenotypes is key to monitor in differentiating cells transcriptional and epigenetic shifts at a functional level and provides a non-genetic means to control cell specification. Expanding the arsenal of analytical tools for metabolic profiling of cell differentiation is therefore of importance. Here, we describe the metabolome of whole pluripotent stem cells (PSCs) using high-resolution magic angle spinning (HR-MAS), a non-destructive approach for Nuclear Magnetic Resonance (NMR) analysis. The integrated H NMR analysis results in detection of metabolites of various groups, including energy metabolites, amino acids, choline derivatives and short chain fatty acids. It unveils new metabolites that discriminate PSCs from differentiated counterparts and directly measures substrates and co-factors of histone modifying enzymes, suggesting that NMR stands as a strategic technique for deciphering metabolic regulations of histone post-translational modifications. HR-MAS NMR analysis of whole PSCs complements the much used solution NMR of cell extracts. Altogether, our multi-platform NMR investigation provides a consolidated picture of PSC metabolic signatures and of metabolic pathways involved in differentiation.
干细胞有望彻底改变当前的医学模式，是监测细胞命运变化的主要“主力军”。在功能水平上，对分化细胞的转录和表观遗传变化进行代谢表型特征分析是关键，它为控制细胞特化提供了一种非遗传手段。因此，拓展细胞分化代谢分析的分析工具组合具有重要意义。在这里，我们使用高分辨率魔角旋转（HR-MAS）技术描述了整个多能干细胞（PSCs）的代谢组，这是一种用于核磁共振（NMR）分析的非破坏性方法。综合 H NMR 分析结果可检测到各种代谢物，包括能量代谢物、氨基酸、胆碱衍生物和短链脂肪酸。它揭示了能够区分 PSCs 和分化细胞的新代谢物，并直接测量组蛋白修饰酶的底物和辅助因子，这表明 NMR 可作为一种解析组蛋白翻译后修饰代谢调控的策略性技术。PSCs 的全细胞 HR-MAS NMR 分析补充了常用于细胞提取物的溶液 NMR。总之，我们的多平台 NMR 研究提供了 PSC 代谢特征以及分化过程中涉及的代谢途径的综合图片。
