Metabolic studies of mammalian cells by 31P-NMR using a continuous perfusion technique

Biochim Biophys Acta. 1984 Jul 20;804(3):275-84. doi: 10.1016/0167-4889(84)90130-7.

Abstract

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.

MeSH terms

  • 2,4-Dinitrophenol
  • Adenosine Triphosphate / metabolism*
  • Animals
  • Cell Line
  • Cricetinae
  • Cricetulus
  • Dinitrophenols / pharmacology
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism*
  • Gels
  • Glucose / pharmacology
  • Lung
  • Magnetic Resonance Spectroscopy
  • Perfusion
  • Phosphates / metabolism*
  • Sodium Chloride / pharmacology

Substances

  • Dinitrophenols
  • Gels
  • Phosphates
  • Sodium Chloride
  • Adenosine Triphosphate
  • Glucose
  • 2,4-Dinitrophenol