Departments of Clinical Research and Radiology, University of Bern, Bern, Switzerland.
Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland and University Institute of Clinical Chemistry, Bern University Hospital, Bern, Switzerland.
Analyst. 2017 Jan 26;142(3):465-471. doi: 10.1039/c6an02195f.
NMR measurements for metabolic characterization of biological samples like cells, biopsies or plasma, may take several hours for advanced methods. Preanalytical issues, such as sample preparation and stability over the measurement time, may have a high impact on metabolite content, and potentially lead to misinterpretation. The aim of this study was therefore to investigate by H HR-MAS NMR the impact of different cell handling preparation protocols on the stability of the cell metabolite content over the measurement time. For this purpose, the metabolite content of fibroblasts and adrenal cells were measured at different time points after lysis and after additional heating. Interestingly the results showed similar metabolite concentrations between lysed and lysed-heated cells at the beginning of the measurement, but increasing differences after some hours of measurement. In lysed cells, metabolism was ongoing, producing metabolite changes over time, contrary to a stable metabolite content of the lysed-heated cells. These results were confirmed in both fibroblasts and adrenal cells. Therefore, in order to minimize metabolite content modifications over the measurement time, it is suggested to use cell lysis in combination with heat inactivation for extended HR-MAS NMR measurements.
NMR 测量可用于对细胞、活检或血浆等生物样本进行代谢特征分析，但对于高级方法可能需要数小时。在分析前，如样品准备和在测量时间内的稳定性等问题，可能会对代谢物含量产生重大影响，并可能导致误解。因此，本研究旨在通过 H HR-MAS NMR 研究不同的细胞处理准备方案对细胞代谢物含量在测量时间内稳定性的影响。为此，在裂解后和加热后不同时间点测量成纤维细胞和肾上腺细胞的代谢物含量。有趣的是，结果表明在测量开始时，裂解和裂解加热细胞之间的代谢物浓度相似，但在测量几个小时后，差异逐渐增大。在裂解细胞中，代谢仍在继续，随着时间的推移产生代谢物变化，而裂解加热细胞的代谢物含量则保持稳定。这一结果在成纤维细胞和肾上腺细胞中均得到了证实。因此，为了尽量减少测量过程中代谢物含量的变化，建议在进行扩展 HR-MAS NMR 测量时，采用细胞裂解与热失活相结合的方法。
