Comprehensive metabolome analysis of intracellular metabolites in cultured cells

Ryuhei Kudo; Kaori Igarashi; Tomoyoshi Soga; Takamasa Ishikawa; Yasuhiro Saito

doi:10.1016/j.xpro.2022.101531

Comprehensive metabolome analysis of intracellular metabolites in cultured cells

STAR Protoc. 2022 Sep 16;3(3):101531. doi: 10.1016/j.xpro.2022.101531. Epub 2022 Jul 11.

Authors

Ryuhei Kudo¹, Kaori Igarashi², Tomoyoshi Soga³, Takamasa Ishikawa⁴, Yasuhiro Saito⁵

Affiliations

¹ Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata 997-0052, Japan; Infinity Lab Co., Ltd., Tsuruoka, Yamagata 997-0052, Japan.
² Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata 997-0052, Japan.
³ Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata 997-0052, Japan. Electronic address: soga@sfc.keio.ac.jp.
⁴ Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata 997-0052, Japan; Infinity Lab Co., Ltd., Tsuruoka, Yamagata 997-0052, Japan. Electronic address: ishikawa@infinity-lab.jp.
⁵ Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata 997-0052, Japan. Electronic address: ysaito@ttck.keio.ac.jp.

Abstract

Capillary electrophoresis mass spectrometry (CE-MS) can measure the intracellular amount of highly polar and charged metabolites; liquid chromatography mass spectrometry (LC-MS) can quantify hydrophobic metabolites. A comprehensive metabolome analysis requires independent sample preparation for LC-MS and CE-MS. Here, we present a protocol to prepare for sequentially analyzing the metabolites from one sample. Here we describe the steps for breast cancer cell lines, MCF-7 cells, but the protocol can be applied to other cell types.

Keywords: Cell Biology; Cell culture; Mass Spectrometry; Metabolism; Metabolomics.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Line
Cells, Cultured
Mass Spectrometry / methods
Metabolome*
Metabolomics* / methods