An understanding of the kinetics of the osmotic response of cells is important in understanding permeability properties of cell membranes and predicting cell responses during exposure to anisotonic conditions. Traditionally, a mathematical model of cell osmotic response is obtained by applying mass transport and Boyle-vant Hoff equations using numerical methods. In the usual application of these equations, it is assumed that all cells are the same size equal to the mean or mode of the population. However, biological cells (even if they had identical membranes and hence identical permeability characteristics--which they do not) have a distribution in cell size and will therefore shrink or swell at different rates when exposed to anisotonic conditions. A population of cells may therefore exhibit a different average osmotic response than that of a single cell. In this study, a mathematical model using mass transport and Boyle-van't Hoff equations was applied to measured size distributions of cells. Chinese hamster fibroblast cells (V-79W) and Madin-Darby canine kidney cells (MDCK), were placed in hypertonic solutions and the kinetics of cell shrinkage were monitored. Consistent with the theoretical predictions, the size distributions of these cells were found to change over time, therefore the selection of the measure of central tendency for the population may affect the calculated osmotic parameters. After examining three different average volumes (mean, median, and mode) using four different theoretical cell size distributions, it was determined that, for the assumptions used in this study, the mean or median were the best measures of central tendency to describe osmotic volume changes in cell suspensions.