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SiMeEx, a simplified method for metabolite extraction of adherent mammalian cells

A reliable method for metabolite extraction is central to mass spectrometry-based metabolomics. However, existing methods are lengthy, mostly due to the step of scraping cells from cell culture vessels, which restricts metabolomics in broader application such as lower cell numbers and high-throughpu...

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Autores principales: Henne, Antonia, Vigh, Anna, Märtens, Andre, Nonnenmacher, Yannic, Ohm, Melanie, Hosseini, Shirin, More, Tushar H., Lauterbach, Mario A., Garritsen, Hendrikus, Korte, Martin, He, Wei, Hiller, Karsten
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9812552/
https://www.ncbi.nlm.nih.gov/pubmed/36619169
http://dx.doi.org/10.3389/fmolb.2022.1084060
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author Henne, Antonia
Vigh, Anna
Märtens, Andre
Nonnenmacher, Yannic
Ohm, Melanie
Hosseini, Shirin
More, Tushar H.
Lauterbach, Mario A.
Garritsen, Hendrikus
Korte, Martin
He, Wei
Hiller, Karsten
author_facet Henne, Antonia
Vigh, Anna
Märtens, Andre
Nonnenmacher, Yannic
Ohm, Melanie
Hosseini, Shirin
More, Tushar H.
Lauterbach, Mario A.
Garritsen, Hendrikus
Korte, Martin
He, Wei
Hiller, Karsten
author_sort Henne, Antonia
collection PubMed
description A reliable method for metabolite extraction is central to mass spectrometry-based metabolomics. However, existing methods are lengthy, mostly due to the step of scraping cells from cell culture vessels, which restricts metabolomics in broader application such as lower cell numbers and high-throughput studies. Here, we present a simplified metabolite extraction (SiMeEx) method, to efficiently and quickly extract metabolites from adherent mammalian cells. Our method excludes the cell scraping step and therefore allows for a more efficient extraction of polar metabolites in less than 30 min per 12-well plate. We demonstrate that SiMeEx achieves the same metabolite recovery as using a standard method containing a scraping step, in various immortalized and primary cells. Omitting cell scraping does not compromise the performance of non-targeted and targeted GC-MS analysis, but enables metabolome analysis of cell culture on smaller well sizes down to 96-well plates. Therefore, SiMeEx demonstrates advantages not only on time and resources, but also on the applicability in high-throughput studies.
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spelling pubmed-98125522023-01-05 SiMeEx, a simplified method for metabolite extraction of adherent mammalian cells Henne, Antonia Vigh, Anna Märtens, Andre Nonnenmacher, Yannic Ohm, Melanie Hosseini, Shirin More, Tushar H. Lauterbach, Mario A. Garritsen, Hendrikus Korte, Martin He, Wei Hiller, Karsten Front Mol Biosci Molecular Biosciences A reliable method for metabolite extraction is central to mass spectrometry-based metabolomics. However, existing methods are lengthy, mostly due to the step of scraping cells from cell culture vessels, which restricts metabolomics in broader application such as lower cell numbers and high-throughput studies. Here, we present a simplified metabolite extraction (SiMeEx) method, to efficiently and quickly extract metabolites from adherent mammalian cells. Our method excludes the cell scraping step and therefore allows for a more efficient extraction of polar metabolites in less than 30 min per 12-well plate. We demonstrate that SiMeEx achieves the same metabolite recovery as using a standard method containing a scraping step, in various immortalized and primary cells. Omitting cell scraping does not compromise the performance of non-targeted and targeted GC-MS analysis, but enables metabolome analysis of cell culture on smaller well sizes down to 96-well plates. Therefore, SiMeEx demonstrates advantages not only on time and resources, but also on the applicability in high-throughput studies. Frontiers Media S.A. 2022-12-21 /pmc/articles/PMC9812552/ /pubmed/36619169 http://dx.doi.org/10.3389/fmolb.2022.1084060 Text en Copyright © 2022 Henne, Vigh, Märtens, Nonnenmacher, Ohm, Hosseini, More, Lauterbach, Garritsen, Korte, He and Hiller. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Molecular Biosciences
Henne, Antonia
Vigh, Anna
Märtens, Andre
Nonnenmacher, Yannic
Ohm, Melanie
Hosseini, Shirin
More, Tushar H.
Lauterbach, Mario A.
Garritsen, Hendrikus
Korte, Martin
He, Wei
Hiller, Karsten
SiMeEx, a simplified method for metabolite extraction of adherent mammalian cells
title SiMeEx, a simplified method for metabolite extraction of adherent mammalian cells
title_full SiMeEx, a simplified method for metabolite extraction of adherent mammalian cells
title_fullStr SiMeEx, a simplified method for metabolite extraction of adherent mammalian cells
title_full_unstemmed SiMeEx, a simplified method for metabolite extraction of adherent mammalian cells
title_short SiMeEx, a simplified method for metabolite extraction of adherent mammalian cells
title_sort simeex, a simplified method for metabolite extraction of adherent mammalian cells
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9812552/
https://www.ncbi.nlm.nih.gov/pubmed/36619169
http://dx.doi.org/10.3389/fmolb.2022.1084060
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