NMR-based metabolomics of mammalian cell and tissue cultures

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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 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.

• Content Type Journal Article
• Pages 1-12
• DOI 10.1007/s10858-011-9490-8
• Authors
  • Nelly Aranibar, Bristol-Myers Squibb Company, Route 206 and Province Line Road, Princeton, NJ 08540, USA
  • Michael Borys, Bristol-Myers Squibb Company, 6000 Thompson Road, Syracuse, NY 13057, USA
  • Nancy A. Mackin, Bristol-Myers Squibb Company, 6000 Thompson Road, Syracuse, NY 13057, USA
  • Van Ly, Bristol-Myers Squibb Company, Route 206 and Province Line Road, Princeton, NJ 08540, USA
  • Nicholas Abu-Absi, Bristol-Myers Squibb Company, 6000 Thompson Road, Syracuse, NY 13057, USA
  • Susan Abu-Absi, Bristol-Myers Squibb Company, 6000 Thompson Road, Syracuse, NY 13057, USA
  • Matthias Niemitz, PERCH Solutions Ltd., Hyrräkatu 3A1, 70500 Kuopio, Finland
  • Bernhard Schilling, Bristol-Myers Squibb Company, 6000 Thompson Road, Syracuse, NY 13057, USA
  • Zheng Jian Li, Bristol-Myers Squibb Company, 6000 Thompson Road, Syracuse, NY 13057, USA
  • Barry Brock, Bristol-Myers Squibb Company, Route 206 and Province Line Road, Princeton, NJ 08540, USA
  • Reb J. Russell, Bristol-Myers Squibb Company, 311 Pennington-Rocky Hill Road, Pennington, NJ 08534, USA
  • Adrienne Tymiak, Bristol-Myers Squibb Company, Route 206 and Province Line Road, Princeton, NJ 08540, USA
  • Michael D. Reily, Bristol-Myers Squibb Company, Route 206 and Province Line Road, Princeton, NJ 08540, USA

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738

Source: Journal of Biomolecular NMR