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题名: High-throughput metabolic profiling based on small amount of hepatic cells
作者: Zhou, Lina1;  Yin, Peiyuan1;  Luo, Ping1;  Tang, Ling1;  Wang, Zhichao1;  Gao, Peng1;  Piao, Hailong1;  Lu, Xin1;  Xu, Guowang1
关键词: A small amount of cells ;  Amino acid metabolism ;  High throughput ;  Metabolic profiling
刊名: ELECTROPHORESIS
发表日期: 2017-09-01
DOI: 10.1002/elps.201600539
卷: 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
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.
关键词[WOS]: EMBRYONIC STEM-CELLS ;  FATTY-ACIDS ;  DRUG DEVELOPMENT ;  CANCER ;  CULTURE ;  CATABOLISM ;  TOXICITY ;  ONCOLOGY ;  GLYCINE ;  DEATH
语种: 英语
WOS记录号: WOS:000410758400007
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/149919
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China

Recommended Citation:
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.
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