Radial Flow Perfusion Enables Real-Time Profiling of Cellular Metabolism at Low Oxygen Levels with Hyperpolarized ¹³C NMR Spectroscopy

• Main Authors: Anthony Mancuso, Mehrdad Pourfathi, Ryan M. Kiefer, Michael C. Noji, Sarmad Siddiqui, Enri Profka, Charles N. Weber, Austin Pantel, Stephen J. Kadlecek, Rahim Rizi, Terence P. F. Gade
• Format: Article
• Language: English
• Published: MDPI AG 2021-08-01
• Series: Metabolites
• Subjects: hyperpolarized ¹³C; DNP; NMR spectroscopy; radial flow; oxygen transport; perfusion
• Online Access: https://www.mdpi.com/2218-1989/11/9/576

In this study, we describe new methods for studying cancer cell metabolism with hyperpolarized ¹³C magnetic resonance spectroscopy (HP ¹³C MRS) that will enable quantitative studies at low oxygen concentrations. Cultured hepatocellular carcinoma cells were grown on the surfaces of non-porous microcarriers inside an NMR spectrometer. They were perfused radially from a central distributer in a modified NMR tube (bioreactor). The oxygen level of the perfusate was continuously monitored and controlled externally. Hyperpolarized substrates were injected continuously into the perfusate stream with a newly designed system that prevented oxygen and temperature perturbations in the bioreactor. Computational and experimental results demonstrated that cell mass oxygen profiles with radial flow were much more uniform than with conventional axial flow. Further, the metabolism of HP [1-¹³C]pyruvate was markedly different between the two flow configurations, demonstrating the importance of avoiding large oxygen gradients in cell perfusion experiments.