Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
Methods Mol Biol. 2025;2855:505-519. doi: 10.1007/978-1-0716-4116-3_27.
Cell cultures are widely used in studies to gain mechanistic insights of metabolic processes. The foundation of these studies lies on the quantification of intracellular and extracellular metabolites, and nuclear magnetic resonance (NMR) is one of the key analytical platforms used to this aim. Among the factors influencing the quality of the produced data are the sampling procedures as well as the acquisition and processing of spectroscopic data. Here we provide our workflow for obtaining quantitative metabolic data from adherent mammalian cells using NMR spectroscopy. The described protocol is compatible with other analytical methods like LC- or GC-MS-based lipidomics and untargeted metabolomics from the same sample. We also show how the collected extracellular data can be used to extract exchange flux rates, particularly useful for flux analysis studies and metabolic engineering of human-induced pluripotent stem cells.
细胞培养广泛应用于研究中，以深入了解代谢过程的机制。这些研究的基础是对细胞内和细胞外代谢物的定量分析，而核磁共振（NMR）是用于实现这一目标的关键分析平台之一。影响所产生数据质量的因素包括采样程序以及光谱数据的采集和处理。在这里，我们提供了一种使用 NMR 光谱法从贴壁哺乳动物细胞中获取定量代谢数据的工作流程。所描述的方案与其他分析方法（如基于 LC 或 GC-MS 的脂质组学和来自同一样本的非靶向代谢组学）兼容。我们还展示了如何利用收集到的细胞外数据提取交换通量率，这对于通量分析研究和人诱导多能干细胞的代谢工程特别有用。
