Metabolic studies of mammalian cells by 31P-NMR using a continuous perfusion technique.
Levels of ATP and Pi in metabolically active Chinese hamster lung fibroblasts were monitored noninvasively by 31P-NMR over many hours and under a variety of conditions. The cells were embedded in a matrix of agarose gel in the form of fine threads which were continuously perfused in a standard NMR tube. The small diameter of the thread allows rapid diffusion of metabolites and drugs into the cells. The changes in ATP and Pi levels were followed as a function of time in response to perfusion with a glucose-containing medium, with isotonic saline and with a medium containing 2,4-dinitrophenol, an uncoupler of oxidative phosphorylation. This gel-thread perfusion method should enable routine NMR studies of cellular metabolism, and may have other potential biological applications.
D019297:2,4-Dinitrophenol; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004140:Dinitrophenols / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005782:Gels; D005947:Glucose / Q000494:pharmacology; D008168:Lung; D009682:Magnetic Resonance Spectroscopy; D010477:Perfusion; D010710:Phosphates / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology
D004140:Dinitrophenols; D005782:Gels; D010710:Phosphates; D012965:Sodium Chloride; D000255:Adenosine Triphosphate; D005947:Glucose; D019297:2,4-Dinitrophenol