NMR-based metabolomics of mammalian cell and tissue cultures.
J Biomol NMR. 2011 Mar 4;
Authors: Aranibar N, Borys M, Mackin NA, Ly V, Abu-Absi N, Abu-Absi S, Niemitz M, Schilling B, Li ZJ, Brock B, Russell RJ, Tymiak A, Reily MD
NMR spectroscopy was used to evaluate growth media and the cellular metabolome in two systems of interest to biomedical research. The first of these was a Chinese hamster ovary cell line engineered to express a recombinant protein. Here, NMR spectroscopy and a quantum mechanical total line shape analysis were utilized to quantify 30 metabolites such as amino acids, Krebs cycle intermediates, activated sugars, cofactors, and others in both media and cell extracts. The impact of bioreactor scale and addition of anti-apoptotic agents to the media on the extracellular and intracellular metabolome indicated changes in metabolic pathways of energy utilization. These results shed light into culture parameters that can be manipulated to optimize growth and protein production. Second, metabolomic analysis was performed on the superfusion media in a common model used for drug metabolism and toxicology studies, in vitro liver slices. In this study, it is demonstrated that two of the 48 standard media components, choline and histidine are depleted at a faster rate than many other nutrients. Augmenting the starting media with extra choline and histidine improves the long-term liver slice viability as measured by higher tissues levels of lactate dehydrogenase (LDH), glutathione and ATP, as well as lower LDH levels in the media at time points out to 94*h after initiation of incubation. In both models, media components and cellular metabolites are measured over time and correlated with currently accepted endpoint measures.
PMID: 21373840 [PubMed - as supplied by publisher]
NMR-based metabolomics of mammalian cell and tissue cultures
NMR-based metabolomics of mammalian cell and tissue cultures
Abstract NMR spectroscopy was used to evaluate growth media and the cellular metabolome in two systems of interest to biomedical research. The first of these was a Chinese hamster ovary cell line engineered to express a recombinant protein. Here, NMR spectroscopy and a quantum mechanical total line shape analysis were utilized to quantify 30 metabolites such as amino acids, Krebs cycle intermediates, activated sugars, cofactors, and others in both media and cell extracts. The impact of bioreactor scale and addition of...
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03-09-2011 04:19 AM
NMR-based stable isotope resolved metabolomics in systems biochemistry
NMR-based stable isotope resolved metabolomics in systems biochemistry
Abstract An important goal of metabolomics is to characterize the changes in metabolic networks in cells or various tissues of an organism in response to external perturbations or pathologies. The profiling of metabolites and their steady state concentrations does not directly provide information regarding the architecture and fluxes through metabolic networks. This requires tracer approaches. NMR is especially powerful as it can be used not only to identify and quantify metabolites in an unfractionated mixture such...
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03-03-2011 02:06 AM
Protocols and applications of cellular metabolomics in safety studies using precision-cut tissue slices and carbon 13 NMR.
Protocols and applications of cellular metabolomics in safety studies using precision-cut tissue slices and carbon 13 NMR.
Protocols and applications of cellular metabolomics in safety studies using precision-cut tissue slices and carbon 13 NMR.
Methods Mol Biol. 2011;691:205-25
Authors: Baverel G, Renault S, Faiz H, El Hage M, Gauthier C, Duplany A, Ferrier B, Martin G
Numerous xenobiotics are toxic to human and animal cells by interacting with their metabolism, but the precise metabolic step affected and the biochemical mechanism behind such a...
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02-08-2011 06:28 PM
Nuclear Magnetic Resonance (NMR)-Based Metabolomics.
Nuclear Magnetic Resonance (NMR)-Based Metabolomics.
Nuclear Magnetic Resonance (NMR)-Based Metabolomics.
Methods Mol Biol. 2011;708:321-34
Authors: Keun HC, Athersuch TJ
Biofluids are by far the most commonly studied sample type in metabolic profiling studies, encompassing blood, urine, cerebrospinal fluid, cell culture media and many others. A number of these fluids can be obtained at a high sampling frequency with minimal invasion, permitting detailed characterisation of dynamic metabolic events. One of the attractive properties of...
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01-06-2011 11:21 AM
[NMR paper] NMR-based metabolomics: a powerful approach for characterizing the effects of environ
NMR-based metabolomics: a powerful approach for characterizing the effects of environmental stressors on organism health.
Related Articles NMR-based metabolomics: a powerful approach for characterizing the effects of environmental stressors on organism health.
Environ Sci Technol. 2003 Nov 1;37(21):4982-9
Authors: Viant MR, Rosenblum ES, Tieerdema RS
It is important to assess the chronic effects of chemical, physical, and biological stressors on organisms in the environment. Appropriate methods must enable rapid, inexpensive, and...