Improving the metabolic stability of cultured cells during extended HR-MAS NMR measurements by prior heating

Diserens, Gaëlle; Hertig, Damian; Eggimann, Sandra; Vermathen, Martina; Furrer, Julien; Nuoffer, Jean-Marc; Vermathen, Peter (27 June 2016). Improving the metabolic stability of cultured cells during extended HR-MAS NMR measurements by prior heating (Unpublished). In: 12th Annual Conference of the Metabolomics Society. Dublin, Ireland. 27.-30.06.2016.

Introduction: 1HHRMAS NMR is an established tool for metabolic characterization of biological samples. However, the accuracy of biomarker detection depends on the sample stability over measurement time. Previously, Duarte et al. found enhanced metabolite visibility and different lipid profiles in lysed compared to frozen cells [1]. Here we investigated effects of shorttime heating prior to HRMAS measurements on metabolite stability of cells. We hypothesized that cell heating has only minor effects on initial metabolite contents (i.e. initially similar spectra with and without heating) and results in increased temporal metabolite stability due to reduced enzymatic activity. Methods: The metabolite content of six lysed nonheated (LFB) and six lysed heated fibroblastsamples (LHFB) was measured as a function of time. Additionally, one sample each of nonheated and heated lysed adrenal cells (LAD & LHAD), which are less robust than fibroblasts, were measured. After lysis according to [1], half of the samples were heated (70°C) for 20min. 1HPROJECT [2] spectra were obtained on a Bruker AvanceII spectrometer (500.13MHz, 277K, MAS=3kHz) at different times over 9 hours. Individual peak analysis was performed investigating temporal changes. For fibroblasts, statistical methods were applied including PCA. Results and Discussion: PCA results and individual peak analysis clearly confirmed our hypothesis: The temporal metabolite stability for LHFB was largely maintained in contrast to LFB. Average temporal variation of all peaks was 14.4% for LHFB compared to 43.1% for LFB. Additionally, the PCA plot demonstrated close clustering for LHFB, whereas LFB were spreading out over time. Also LHAD showed temporal metabolic stability, while LAD exhibited strong changes. Our results suggest using cell lysis in combination with heating for extended longterm HRMAS NMR measurements, in order to minimize metabolite content modifications over measurement time. References: 1. Duarte IF, et al. Anal. Chem. 2009;81:5023. 2. Aguilar JA, et al. Chemical Communications 2012;48:811.

Item Type:

Conference or Workshop Item (Poster)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry
04 Faculty of Medicine > Department of Radiology, Neuroradiology and Nuclear Medicine (DRNN) > Institute of Diagnostic, Interventional and Paediatric Radiology > DCR Magnetic Resonance Spectroscopy and Methodology (AMSM)
04 Faculty of Medicine > Department of Haematology, Oncology, Infectious Diseases, Laboratory Medicine and Hospital Pharmacy (DOLS) > Institute of Clinical Chemistry
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Pavillon 52 > Abt. Magnetresonanz-Spektroskopie und Methodologie, AMSM

UniBE Contributor:

Diserens, Gaëlle; Vermathen, Martina; Furrer, Julien; Nuoffer, Jean-Marc and Vermathen, Peter

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry
600 Technology > 610 Medicine & health

Funders:

[4] Swiss National Science Foundation

Language:

English

Submitter:

Martina Vermathen

Date Deposited:

26 Jan 2017 09:06

Last Modified:

26 Jan 2017 09:06

URI:

https://boris.unibe.ch/id/eprint/92517

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