Fast filtration for metabolome sampling of suspended animal cells
Fast filtration for metabolome sampling of suspended animal cells
复制标题
用于悬浮动物细胞代谢组采样的快速过滤
DOI：
10.1007/s10529-010-0466-7
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发表时间：
2011
影响因子：
2.7
通讯作者：
T. Noll
中科院分区：
文献类型：
--
作者：
M. Volmer;Stefan Northoff;S. Scholz;T. Thüte;H. Büntemeyer;T. Noll
本文提出了一种快速过滤取样悬浮动物细胞的新方法，该方法能快速淬灭细胞代谢，并能有效地从培养基中分离细胞。与用微结构热交换器或离心而没有预先淬灭的采样相比，腺苷酸能荷和测量的浓度，特别是具有高周转率的代谢物或代谢途径早期的代谢物的腺苷酸能荷和测量的浓度显著较高。当在洗涤步骤中使用等渗NaCl溶液时，没有观察到ATP从细胞中泄漏。因此，快速过滤和冷甲醇提取的组合适用于悬浮动物细胞培养物的细胞内代谢组学研究，并且优于目前应用的其他方法上级。
A new method for sampling suspended animal cells by fast filtration is presented that allows rapid quenching of cellular metabolism and efficient separation of the cells from culture medium. Compared to sampling with a microstructure heat exchanger or centrifugation without prior quenching, the adenylate energy charge and the measured concentrations especially of metabolites with a high turnover rate or of metabolites early in metabolic pathways were substantially higher. No leakage of ATP from the cells was observed when using iso-osmotic NaCl solution in the washing step. The combination of fast filtration and cold methanol extraction is therefore suitable for intracellular metabolomic studies of suspended animal cell cultures and superior to other methods currently applied.
影响因子：
7.4
作者：
Sellick, Christopher A.;Hansen, Rasmus;Dickson, Alan J.
通讯作者：
Dickson, Alan J.