Removing the bottlenecks of cell culture metabolomics: fast normalization procedure, correlation of metabolites to cell number, and impact of the cell harvesting method.
Removing the bottlenecks of cell culture metabolomics: fast normalization procedure, correlation of metabolites to cell number, and impact of the cell harvesting method.
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DOI：
10.1007/s11306-016-1104-8
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发表时间：
2016
期刊：
影响因子：
3.6
通讯作者：
Tokarz, Janina
中科院分区：
文献类型：
--
作者：
Muschet, Caroline;Moeller, Gabriele;Prehn, Cornelia;de Angelis, Martin Hrabe;Adamski, Jerzy;Tokarz, Janina
尽管目前已有代谢组学技术定期对培养细胞进行分析，但研究设置和数据处理中的一些问题仍未得到完全解决：贴壁细胞的合适采集方法，数据归一化的快速和稳健方法，以及代谢物水平可归一化为细胞数量的证据。我们打算开发一种细胞培养代谢组学样本标准化的快速方法，分析代谢物水平如何与细胞数量相关，并阐明收获类型对所测量代谢物图谱的影响。我们培养了四种不同的人类细胞系，并利用它们开发了一种基于荧光的DNA定量方法。此外，我们评估了代谢物水平和细胞数量之间的相关性，并重点研究了采集方法(刮除或胰酶消化)对代谢物图谱的影响。我们开发了一种快速、灵敏和稳健的基于荧光的DNA定量方法，显示了所有细胞系的荧光强度与细胞数量之间良好的线性相关性。此外，82-97%的细胞内代谢物浓度与细胞数量呈线性相关。我们观察到胰酶消化的细胞和刮除的细胞在氨基酸、生物胺和脂肪水平上的差异。我们为细胞培养代谢组学样本提供了一种快速、稳健和有效的归一化方法，并证明了代谢组学数据归一化符合细胞数量的条件。我们展示了细胞系和代谢物特定的收获方法对代谢物浓度的影响。本文的在线版本(doi：10.1007/s11306-016-1104-8)包含补充材料，授权用户可以使用。
Although cultured cells are nowadays regularly analyzed by metabolomics technologies, some issues in study setup and data processing are still not resolved to complete satisfaction: a suitable harvesting method for adherent cells, a fast and robust method for data normalization, and the proof that metabolite levels can be normalized to cell number. We intended to develop a fast method for normalization of cell culture metabolomics samples, to analyze how metabolite levels correlate with cell numbers, and to elucidate the impact of the kind of harvesting on measured metabolite profiles. We cultured four different human cell lines and used them to develop a fluorescence-based method for DNA quantification. Further, we assessed the correlation between metabolite levels and cell numbers and focused on the impact of the harvesting method (scraping or trypsinization) on the metabolite profile. We developed a fast, sensitive and robust fluorescence-based method for DNA quantification showing excellent linear correlation between fluorescence intensities and cell numbers for all cell lines. Furthermore, 82–97 % of the measured intracellular metabolites displayed linear correlation between metabolite concentrations and cell numbers. We observed differences in amino acids, biogenic amines, and lipid levels between trypsinized and scraped cells. We offer a fast, robust, and validated normalization method for cell culture metabolomics samples and demonstrate the eligibility of the normalization of metabolomics data to the cell number. We show a cell line and metabolite-specific impact of the harvesting method on metabolite concentrations. The online version of this article (doi:10.1007/s11306-016-1104-8) contains supplementary material, which is available to authorized users.
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影响因子：
3.6
作者：
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