Title: Hyperpolarized (13)C spectroscopy and an NMR-compatible bioreactor system for the investigation of real-time cellular metabolism.

Authors: Keshari, Kayvan R; Kurhanewicz, John; Jeffries, Rex E; Wilson, David M; Dewar, Brian J; Van Criekinge, Mark; Zierhut, Matthew; Vigneron, Daniel B; Macdonald, Jeffrey M

Published In Magn Reson Med, (2010 Feb)

Abstract: The purpose of this study was to combine a three-dimensional NMR-compatible bioreactor with hyperpolarized (13)C NMR spectroscopy in order to probe cellular metabolism in real time. JM1 (immortalized rat hepatoma) cells were cultured in a three-dimensional NMR-compatible fluidized bioreactor. (31)P spectra were acquired before and after each injection of hyperpolarized [1-(13)C] pyruvate and subsequent (13)C spectroscopy at 11.7 T. (1)H and two-dimensional (1)H-(1)H-total correlation spectroscopy spectra were acquired from extracts of cells grown in uniformly labeled (13)C-glucose, on a 16.4 T, to determine (13)C fractional enrichment and distribution of (13)C label. JM1 cells were found to have a high rate of aerobic glycolysis in both two-dimensional culture and in the bioreactor, with 85% of the (13)C label from uniformly labeled (13)C-glucose being present as either lactate or alanine after 23 h. Flux measurements of pyruvate through lactate dehydrogenase and alanine aminotransferase in the bioreactor system were 12.18 +/- 0.49 nmols/sec/10(8) cells and 2.39 +/- 0.30 nmols/sec/10(8) cells, respectively, were reproducible in the same bioreactor, and were not significantly different over the course of 2 days. Although this preliminary study involved immortalized cells, this combination of technologies can be extended to the real-time metabolic exploration of primary benign and cancerous cells and tissues prior to and after therapy.

PubMed ID: 20099325

MeSH Terms: No MeSH terms associated with this publication