Henne Antonia, Vigh Anna, Märtens Andre, Nonnenmacher Yannic, Ohm Melanie, Hosseini Shirin, More Tushar H, Lauterbach Mario A, Garritsen Hendrikus, Korte Martin, He Wei, Hiller Karsten
Department of Bioinformatics and Biochemistry, Braunschweig Integrated Centre of Systems Biology, Technische Universität Braunschweig, Braunschweig, Germany.
Department of Cellular Neurobiology, Zoological Institute, Technische Universität Braunschweig, Braunschweig, Germany.
Front Mol Biosci. 2022 Dec 21;9:1084060. doi: 10.3389/fmolb.2022.1084060. eCollection 2022.
A reliable method for metabolite extraction is central to mass spectrometry-based metabolomics. However, existing methods are lengthy, mostly due to the step of scraping cells from cell culture vessels, which restricts metabolomics in broader application such as lower cell numbers and high-throughput studies. Here, we present a simplified metabolite extraction (SiMeEx) method, to efficiently and quickly extract metabolites from adherent mammalian cells. Our method excludes the cell scraping step and therefore allows for a more efficient extraction of polar metabolites in less than 30 min per 12-well plate. We demonstrate that SiMeEx achieves the same metabolite recovery as using a standard method containing a scraping step, in various immortalized and primary cells. Omitting cell scraping does not compromise the performance of non-targeted and targeted GC-MS analysis, but enables metabolome analysis of cell culture on smaller well sizes down to 96-well plates. Therefore, SiMeEx demonstrates advantages not only on time and resources, but also on the applicability in high-throughput studies.
一种可靠的代谢物提取方法是基于质谱的代谢组学的核心。然而,现有方法耗时较长,主要是因为从细胞培养容器中刮取细胞这一步骤,这限制了代谢组学在细胞数量较少和高通量研究等更广泛应用中的开展。在此,我们提出了一种简化代谢物提取(SiMeEx)方法,以高效、快速地从贴壁哺乳动物细胞中提取代谢物。我们的方法省去了细胞刮取步骤,因此每12孔板能在不到30分钟的时间内更高效地提取极性代谢物。我们证明,在各种永生化细胞和原代细胞中,SiMeEx与使用包含刮取步骤的标准方法具有相同的代谢物回收率。省去细胞刮取步骤不会影响非靶向和靶向气相色谱 - 质谱分析的性能,反而能够对小至96孔板的较小孔尺寸的细胞培养物进行代谢组分析。因此,SiMeEx不仅在时间和资源方面具有优势,在高通量研究中的适用性也具有优势。