Hyperpolarized 13 C spectroscopy and an NMR‐compatible bioreactor system for the investigation of real‐time cellular metabolism

• Published in: Magnetic resonance in medicine Vol. 63; no. 2; pp. 322 - 329
• Main 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.
• Format: Journal Article
• Language: English
• Published: 01-02-2010
• ISSN: 0740-3194; 1522-2594
• DOI: 10.1002/mrm.22225

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. Magn Reson Med, 2010. © 2010 Wiley‐Liss, Inc.