Effects of cell density on in vitro glucose metabolism by isolated adipocytes

We studied the effect of variable isolated fat cell concentrations (from 0.17 to 1.25 x 10⁶ cells/ml) on rate and pattern of basal and insulin- stimulated glucose metabolism by rat epididymal fat cells. Cell concentration did not affect total glucose utilization, but high cell concentrations increased the absolute and relative conversion of glucose to CO₂ and glyceride-fatty acids by two- to threefold and decreased the conversion to lactate, pyruvate, and glyceride-glycerol when compared with values observed at low cell concentration. When effects of adenosine deaminase (ADA) and N- 6(2-phenylisopropyl)adenosine (PIA) were examined, addition of ADA to incubated cells produced no significant changes in the rate or pattern of adipocyte glucose metabolism; PIA had a slight and uniform effect on the conversion of glucose to its metabolic products and minimal effect on insulin-stimulated glucose metabolism. Medium free fatty acid concentration did not change during the incubation at various cell density, but intracellular free fatty acids were found to be inversely related to fat cell density in the medium. Thus a variable fat cell density influences the pattern of adipocyte glucose metabolism in vitro. This effect may be due to variable rates of lipolysis and resulting changes in intracellular fatty acid concentration rather than to adenosine per se. This work has practical implications in the need to define cell density when carrying out in vitro measurements of adipocyte glucose conversion to products.