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Measurement by Nuclear Magnetic Resonance Diffusion of the Dimensions of the Mobile Lipid Compartment in C6 Cells¹

Unitat de Ciències, Departament de Bioquímica i Biologia Molecular [Y. P., M. E. C., C. A.], Servei de Ressonància Magnètica Nuclear [M. E. C.], and Unitat de Biofísica de Medicina, Departament de Bioquímica i Biologia Molecular [R. B., M. S.], Universitat Autònoma de Barcelona, 08193 Cerdanyola del Vallès, Spain, and Unité Mixte Université Joseph Fourier-INSERM U438, Laboratoire de Recherche Correspondant du Commissariut a l’Energie Atomique, Centre Hospitalier Universitaire de Grenoble, BP 217, 38043 Grenoble Cedex 9, France [H. L., C. R.]

The ¹H spectrum of certain tumor cells, in vivo tumors, and their biopsies in vitro shows a narrow and intense resonance at 1.26 ppm, which has been assigned to the fatty acyl chain of triglycerides [nuclear magnetic resonance (NMR) visible mobile lipids, MLs]. We have used diffusion-weighted NMR spectroscopy to directly address the subcellular origin of MLs in the case of C6 cells in which lactate accumulation had been inhibited by prior iodoacetamide incubation. Borage oil and artificial lipid droplets were used as model systems of free and restricted diffusion, respectively. The characteristic diameter for the ML resonance compartment measured by NMR for the C6 cells was not significantly different from the one obtained with phase contrast microscopy (1.88 ± 0.04 µm from NMR versus 1.37 ± 0.33 µm from microscopy). We herewith provide direct and noninvasive evidence that the lipid signal at 1.26 ppm in C6 cells, which remains visible in long echo time (T_E = 136 ms) experiments, mostly originates from subcellular structures with diameters of 1–2 µm, which correspond to the cytosolic lipid droplets that can be detected in optical microscopy preparations of the same cells.