A method using 3-O-methyl-D-glucose and phloretin for the determination of intracellular water space of cells in monolayer culture.

Abstract A technique is described by which the intracellular water space can be determined in cells which are attached to a culture dish. A major advantage of this procedure is that the cells maintain their normal morphology. This technique takes advantage of the transport of the nonmetabolizable hexose, 3- O -methyl- d -glucose, to an intracellular concentration equal to the extracellular concentration, and the inhibition of 3- O -methyl- d -glucose efflux by phloretin. It requires only a determination of the nanomoles of hexose taken up at equilibrium plus a protein or cell number determination.