In vitro detection of small molecule metabolites excreted from cancer cells using a Tenax TA thin-film microextraction device.
We developed a new device for the in vitro extraction of small molecule metabolites excreted from cancer cells. The extraction device, which was biocompatible and incubated with cancer cells, consists of a thin Tenax TA film deposited on the surface of a cylindrical aluminum rod. The Tenax TA solid phase was utilized for the direct extraction and preconcentration of the small molecule metabolites from a cell culture sample. The device fabrication and the metabolite extraction were optimized, tested, and validated using HeLa cell cultures. Comparison of metabolic profiles with the control measurement from the culture medium enabled detection of metabolites that were consumed or produced by the cell culture. Tentative identification and semi-quantitative investigation of the excreted metabolites were performed by GC-MS analysis. The proposed approach can be a valuable tool for the characterization of low-volatile cancer cell metabolites that are not covered by use of conventional methods based on headspace solid phase microextraction.
Graduate School of Environmental Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu, Kitakyushu 808-0135, Japan.
Takuma
T
Nozoe
Graduate School of Environmental Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu, Kitakyushu 808-0135, Japan.
Shigemi
S
Goda
Graduate School of Environmental Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu, Kitakyushu 808-0135, Japan; WPI International Institute for Carbon-Neutral Energy Research (WPI-I(2) CNER), Kyushu University, Fukuoka 819-0395, Japan.
Roman
R
Selyanchyn
Graduate School of Environmental Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu, Kitakyushu 808-0135, Japan.
Tao
T
Wang
Graduate School of Environmental Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu, Kitakyushu 808-0135, Japan.
Kohji
K
Nakazawa
Graduate School of Environmental Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu, Kitakyushu 808-0135, Japan.
Takeshi
T
Hirano
Shinkou Seiki Co. Ltd., Fukuoka 812-0054, Japan.
Hidetaka
H
Matsui
Graduate School of Environmental Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu, Kitakyushu 808-0135, Japan. Electronic address: leesw@kitakyu-u.ac.jp.
Seung-Woo
SW
Lee
D018929:Cell Culture Techniques; D004867:Equipment Design; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D009369:Neoplasms / Q000737:chemistry*; D011108:Polymers; D052617:Solid Phase Microextraction / Q000295:instrumentation* / Q000379:methods; D055549:Volatile Organic Compounds / Q000032:analysis / Q000302:isolation & purification*
D011108:Polymers; D055549:Volatile Organic Compounds; C010514:tenax
Cell culture; GC–MS; Molecular cancer biomarkers; Thin-film microextraction