Impact of sample preparation upon intracellular metabolite measurements in 3D cell culture systems.

Détails

Ressource 1Télécharger: 31190156.pdf (1507.11 [Ko])
Etat: Public
Version: Final published version
Licence: CC BY 4.0
ID Serval
serval:BIB_56A04C793F67
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Impact of sample preparation upon intracellular metabolite measurements in 3D cell culture systems.
Périodique
Metabolomics
Auteur(s)
Mathon C., Bovard D., Dutertre Q., Sendyk S., Bentley M., Hoeng J., Knorr A.
ISSN
1573-3890 (Electronic)
ISSN-L
1573-3882
Statut éditorial
Publié
Date de publication
12/06/2019
Peer-reviewed
Oui
Volume
15
Numéro
6
Pages
92
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Résumé
Interest in cell culture metabolomics has increased greatly in recent years because of its many potential applications and advantages (e.g., in toxicology). The first critical step for exploring the cellular metabolome is sample preparation. For metabolomics studies, an ideal sample preparation would extract a maximum number of metabolites and would enable reproducible, accurate analysis of a large number of samples and replicates. In addition, it would provide consistent results across several studies over a relatively long time frame.
This study was conducted to evaluate the impact of sample preparation strategies on monitoring intracellular metabolite responses, highlighting the potential critical step(s) in order to finally improve the quality of metabolomics studies.
The sample preparation strategies were evaluated by calculating the sample preparation effect, matrix factor, and process efficiency (PE) for 16 tobacco exposition-related metabolites, including nicotine, nicotine-derived nitrosamine ketone, their major metabolites, and glutathione, using isotopically-labelled internal standards. Samples were analyzed by liquid chromatography (LC) coupled to high-resolution mass spectrometry (HRMS).
A sample drying step increased losses or variability for some selected metabolites. By avoiding evaporation, good sample preparation recovery was obtained for these compounds. For some metabolites, the cell or culture type impacted PE and matrix factor.
In our sample preparation protocol, the drying-reconstitution step was identified as the main cause of metabolite losses or increased data variability during metabolomics analysis by LC-HRMS. Furthermore, PE was affected by the type of matrix. Isotopologue internal standards fully compensate losses or enhancements.
Mots-clé
Bronchi/cytology, Bronchi/metabolism, Cell Culture Techniques/methods, Cell Line, Chemical Fractionation/methods, Chromatography, Liquid/methods, Epithelial Cells/cytology, Epithelial Cells/metabolism, Glutathione/metabolism, Humans, Mass Spectrometry/methods, Metabolome, Metabolomics/methods, Nicotine/metabolism, Spheroids, Cellular/cytology, Spheroids, Cellular/metabolism, Cell culture, Effectiveness, Extraction, Intracellular metabolites, LC-HRMS, Sample preparation
Pubmed
Web of science
Open Access
Oui
Création de la notice
19/06/2020 10:12
Dernière modification de la notice
15/05/2021 6:09
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