Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń, Toruń, Poland.
Department of Chemistry, Middle East Technical University, Ankara, 06800, Turkey.
Sci Rep. 2019 Jan 23;9(1):402. doi: 10.1038/s41598-018-36481-2.
Use of solid phase microextraction (SPME) for cell culture metabolomic analysis allows for the attainment of more sophisticated data from in vitro cell cultures. Moreover, considering that SPME allows the implementation of multiple extractions from the same sample due to its non/low-depletive nature, time course studies using the same set of samples are thus facilitated via this method. Such an approach results in a reduction in the number of samples needed for analysis thus eliminates inter-batch variability related to biological variation occurring during cell culturing. The current work aims to demonstrate the capability of SPME for measurements of combretastatin A4 (CA4) effectiveness on non-small cell cancer cell line. A cultivation protocol was established in the 96-well plate, and a fiber format of SPME was selected for metabolite extraction. The extracellular metabolic pattern of cells was changed after administration of the tested drug. This suggests pharmacological activity of the administered compound towards the studied cell line model. Results support that the use of direct immersion SPME for analysis of cell cultures does not affect cells growth or contaminate sample. Consequently, SPME allows the attainment of accurate information regarding drug uptake, metabolism, and metabolomic changes in the studied cells induced by exposure to the drug simultaneously in a single experiment.
固相微萃取(SPME)在细胞培养代谢组学分析中的应用可实现更复杂的体外细胞培养数据。此外,由于 SPME 具有非/低耗竭性,允许从同一样品中进行多次提取,因此通过这种方法可以方便地进行相同样品集的时间过程研究。这种方法减少了分析所需的样品数量,从而消除了与细胞培养过程中发生的生物学变异相关的批次间变异性。本工作旨在展示 SPME 测量 combretastatin A4(CA4)对非小细胞癌细胞系有效性的能力。建立了 96 孔板的培养方案,并选择纤维格式的 SPME 进行代谢物提取。给药后细胞的细胞外代谢模式发生了变化。这表明所测试的化合物对所研究的细胞系模型具有药理学活性。结果支持直接浸入式 SPME 用于细胞培养分析不会影响细胞生长或污染样品。因此,SPME 允许在单个实验中同时获得关于药物摄取、代谢和暴露于药物引起的研究细胞代谢组变化的准确信息。