title: Metabolic stability of cells for extended metabolomical measurements using NMR. A comparison between lysed and additionally heat inactivated cells.
abstract: NMR measurements for metabolic characterization of biological samples like cells, biopsies or plasma, may take several hours for advanced methods. Preanalytical issues, such as sample preparation and stability over the measurement time, may have a high impact on metabolite content, and potentially lead to misinterpretation. The aim of this study was therefore to investigate by1H HR-MAS NMR the impact of different cell handling preparation protocols on the stability of the cell metabolite content over the measurement time. For this purpose, the metabolite content of fibroblasts and adrenal cells were measured at different time points after lysis and after additional heating. Interestingly the results showed similar metabolite concentrations between lysed and lysed-heated cells at the beginning of the measurement, but increasing differences after some hours of measurement. In lysed cells, metabolism was ongoing, producing metabolite changes over time, contrary to a stable metabolite content of the lysed-heated cells. These results were confirmed in both fibroblasts and adrenal cells. Therefore, in order to minimize metabolite content modifications over the measurement time, it is suggested to use cell lysis in combination with heat inactivation for extended HR-MAS NMR measurements.
authors.affiliation: Departments of Clinical Research and Radiology, University of Bern, Bern, Switzerland. gaelle.diserens@insel.ch.
authors.forename: G
authors.identifier: http://orcid.org/0000-0002-6153-4888
authors.initials: G
authors.lastname: Diserens
authors.affiliation: Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland and University Institute of Clinical Chemistry, Bern University Hospital, Bern, Switzerland.
authors.forename: D
authors.initials: D
authors.lastname: Hertig
authors.affiliation: Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Vermathen
authors.affiliation: Pediatric Endocrinology and Diabetology of the Department of Pediatrics and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: B
authors.initials: B
authors.lastname: Legeza
authors.affiliation: Pediatric Endocrinology and Diabetology of the Department of Pediatrics and Department of Clinical Research, University of Bern, Bern, Switzerland.
authors.forename: C E
authors.initials: CE
authors.lastname: Flück
authors.affiliation: University Institute of Clinical Chemistry, Bern University Hospital, Bern, Switzerland.
authors.forename: J M
authors.initials: JM
authors.lastname: Nuoffer
authors.affiliation: Departments of Clinical Research and Radiology, University of Bern, Bern, Switzerland. gaelle.diserens@insel.ch.
authors.forename: P
authors.initials: P
authors.lastname: Vermathen
mesh_terms: D000311:Adrenal Glands / Q000166:cytology*; D001706:Biopsy; D002460:Cell Line; D005347:Fibroblasts / Q000166:cytology*; D006358:Hot Temperature; D006801:Humans; D055442:Metabolome; D066244:Proton Magnetic Resonance Spectroscopy; D012867:Skin / Q000166:cytology