Hyperpolarized (13)C spectroscopy and an NMR-compatible bioreactor system for the investigation of real-time cellular metabolism.
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.
Keshari
Kayvan R
KR
Department of Radiology, University of California San Francisco, SanFrancisco, California, USA.
Kurhanewicz
John
J
Jeffries
Rex E
RE
Wilson
David M
DM
Dewar
Brian J
BJ
Van Criekinge
Mark
M
Zierhut
Matthew
M
Vigneron
Daniel B
DB
Macdonald
Jeffrey M
JM
D000818:Animals; D019149:Bioreactors; D002247:Carbon Isotopes / Q000032:analysis; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000295:instrumentation*; D045744:Cell Line, Tumor; D004867:Equipment Design; D019544:Equipment Failure Analysis; D009682:Magnetic Resonance Spectroscopy / Q000295:instrumentation*; D009363:Neoplasm Proteins / Q000032:analysis*; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D002247:Carbon Isotopes; D009363:Neoplasm Proteins