In Vitro Liquid Extraction Surface Analysis Mass Spectrometry (ivLESA-MS) for Direct Metabolic Analysis of Adherent Cells in Culture.
In Vitro Liquid Extraction Surface Analysis Mass Spectrometry (ivLESA-MS) for Direct Metabolic Analysis of Adherent Cells in Culture.
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DOI：
10.1021/acs.analchem.8b00530
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发表时间：
2018-04-17
影响因子：
7.4
通讯作者：
Agar NYR
中科院分区：
文献类型：
--
作者：
Basu SS;Randall EC;Regan MS;Lopez BGC;Clark AR;Schmitt ND;Agar JN;Dillon DA;Agar NYR
传统的代谢组学方法包括大量的样品制备步骤和较长的分析运行时间，增加了处理伪影的可能性并限制了高通量应用。我们在这里提出了体外液体萃取表面分析质谱（ivLESA-MS），LESA-MS的变化，直接在96孔细胞培养板中生长的贴壁细胞上进行。为了实现这一点，在分析前立即抽吸培养基，并使用LESA从细胞单层表面提取代谢物，然后以负离子模式进行纳米电喷雾电离和MS分析。我们应用该平台来表征和比较在培养物中生长的多种乳腺癌细胞系（MCF-7、ZR-75-1、MDA-MB-453和MDA-MB-231）的脂质组学特征，并揭示了细胞系之间不同且可重复的脂质组学特征。此外，我们还展示了依赖于时间的处理工件，强调了即时分析的重要性。ivLESA-MS代表了一种快速的体外代谢组学方法，该方法无需淬灭、细胞收获、样品制备和色谱法，显著缩短了制备和分析时间，同时最大限度地减少了处理伪影。该方法可进一步适用于以高通量方式在体外测试药物。
Conventional metabolomic methods include extensive sample preparation steps and long analytical run times, increasing the likelihood of processing artifacts and limiting high throughput applications. We present here in vitro liquid extraction surface analysis mass spectrometry (ivLESA-MS), a variation on LESA-MS, performed directly on adherent cells grown in 96-well cell culture plates. To accomplish this, culture medium was aspirated immediately prior to analysis, and metabolites were extracted using LESA from the cell monolayer surface, followed by nano-electrospray ionization and MS analysis in negative ion mode. We applied this platform to characterize and compare lipidomic profiles of multiple breast cancer cell lines growing in culture (MCF-7, ZR-75–1, MDA-MB-453, and MDA-MB-231) and revealed distinct and reproducible lipidomic signatures between the cell lines. Additionally, we demonstrated time-dependent processing artifacts, underscoring the importance of immediate analysis. ivLESA-MS represents a rapid in vitro metabolomic method, which precludes the need for quenching, cell harvesting, sample preparation, and chromatography, significantly shortening preparation and analysis time while minimizing processing artifacts. This method could be further adapted to test drugs in vitro in a high throughput manner.
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DOI：
10.1073/pnas.1408129111
发表时间：
2014-10-21
影响因子：
11.1
作者：
Calligaris, David;Caragacianu, Diana;Agar, Nathalie Y. R.
通讯作者：
Agar, Nathalie Y. R.
影响因子：
5.3
作者：
Huang WC;Chen CL;Lin YS;Lin CF
通讯作者：
Lin CF
DOI：
10.1073/pnas.1404724111
发表时间：
2014-07-29
影响因子：
11.1
作者：
Santagata, Sandro;Eberlin, Livia S.;Agar, Nathalie Y. R.
通讯作者：
Agar, Nathalie Y. R.
影响因子：
7.4
作者：
Dethy, JM;Ackermann, BL;Schultz, GA
通讯作者：
Schultz, GA
影响因子：
4.8
作者：
London, M;London, E
通讯作者：
London, E