Hyperpolarized [1-13C] α-KG Metabolism

A volume of 30μL of [1-¹³C] α-KG solution (5.9M, 3:1 water:glycerol, 17.3mM OX63 radical, 0.4mM Dotarem) was polarized using a hypersense polarizer (Oxford Instruments) for approximately 1 hour (34 (link)). Hyperpolarized [1-¹³C] α-KG was then rapidly dissolved in isotonic buffer (40mM Tris, 30mM NaOH, 3.0μmol/L Na₂EDTA) and injected into the perfusion medium of U87IDHwt cells (n=5) and U87IDHmut cells (n=5) to a final concentration of 15mM. Dynamic sets of hyperpolarized ¹³C spectra were acquired on a 11.7 Tesla INOVA spectrometer (Agilent Technologies, Inc., Palo Alto, CA) starting at the beginning of the injection using a pulse-acquire sequence (13 degrees flip angle (FA), 3 second repetition time (TR), acquisition time 300 seconds)(25 (link)).