Continuous measurement of glucose utilization in heart myoblasts.
Understanding quantitative aspects of cell energy metabolism and how it is influenced by environment is central to biology, medicine, and biotechnology. Most methods used for measuring metabolic fluxes associated with energy metabolism require considerable personnel effort or high maintenance instrumentation. The microphysiometer is a commercially available instrument that measures acid extrusion rates, which are commonly used for drug screening. With the addition of oxygen sensors, the instrument can also be used to measure cell oxygen consumption rates and thereby calculate glycolytic fluxes. In the work described here, oxygen consumption and acid extrusion rates were used to measure glucose utilization by the H9c2 rat heart myoblast cell line and these results are compared with fluxes measured with a radiometric assay. Both assays were used to investigate changes in H9c2 energy metabolism due to cell stimulation with carbachol and insulin. The results demonstrate the utility of the microphysiometer method for measuring both transient and sustained changes in partitioning of glucose utilization between glycolysis and oxidation in live cells.
Wiley
Carmen
C
Department of Chemistry, University of Washington, Seattle, WA 98195-1700, USA.
Beeson
Craig
C
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation; D002460:Cell Line; D004734:Energy Metabolism / Q000502:physiology; D005947:Glucose / Q000378:metabolism*; D006019:Glycolysis / Q000502:physiology; D019344:Lactic Acid / Q000378:metabolism; D016393:Lymphoma, B-Cell / Q000378:metabolism; D051379:Mice; D015336:Molecular Probe Techniques; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D010101:Oxygen Consumption / Q000502:physiology; D011874:Radiometry; D051381:Rats; D014407:Tumor Cells, Cultured
D019344:Lactic Acid; D005947:Glucose