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High-throughput metabolic profiling based on small amount of hepatic cells
Zhou, Lina; Yin, Peiyuan; Luo, Ping; Tang, Ling; Wang, Zhichao; Gao, Peng; Piao, Hailong; Lu, Xin; Xu, Guowang
关键词a Small Amount Of Cells Amino Acid Metabolism High Throughput Metabolic Profiling
刊名ELECTROPHORESIS
2017-09-01
DOI10.1002/elps.201600539
38期:18页:2296-2303
收录类别SCI
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences
类目[WOS]Biochemical Research Methods ; Chemistry, Analytical
研究领域[WOS]Biochemistry & Molecular Biology ; Chemistry
关键词[WOS]EMBRYONIC STEM-CELLS ; FATTY-ACIDS ; DRUG DEVELOPMENT ; CANCER ; CULTURE ; CATABOLISM ; TOXICITY ; ONCOLOGY ; GLYCINE ; DEATH
英文摘要Common metabolomics platforms require about 10(6) cells, which has a limited throughput due to the time-consuming steps of cell culture and preparation. There is a demand for metabolic profiling methods to improve analytical throughput and detection sensitivity based on small amount of cells. In this study, we proposed a high-throughput scheme, integrating 96-well plate cell cultivation, in-situ cell pretreatment, and sensitive dansylation labeling coupled with LC-MS analysis of metabolites inside HepG2 cells (of the order of magnitude of 10(3) cells in each well). A simple and rapid cell pretreatment was performed showing good extraction efficiency and good precision (the RSDs smaller than 5%) for polar metabolites. The recovery in metabolite extraction evaluated with three isotope-labeled amino acids was from 89.7 to 106.3% at low, medium, and high concentrations. The suitability of the method was illustrated by exploring influences of different fatty acids on HepG2 cells.
语种英语
WOS记录号WOS:000410758400007
引用统计
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/149919
专题中国科学院大连化学物理研究所
作者单位Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
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Zhou, Lina,Yin, Peiyuan,Luo, Ping,et al. High-throughput metabolic profiling based on small amount of hepatic cells[J]. ELECTROPHORESIS,2017,38(18):2296-2303.
APA Zhou, Lina.,Yin, Peiyuan.,Luo, Ping.,Tang, Ling.,Wang, Zhichao.,...&Xu, Guowang.(2017).High-throughput metabolic profiling based on small amount of hepatic cells.ELECTROPHORESIS,38(18),2296-2303.
MLA Zhou, Lina,et al."High-throughput metabolic profiling based on small amount of hepatic cells".ELECTROPHORESIS 38.18(2017):2296-2303.
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