Osmotic volume change and cellular injury by hypertonic sodium choloride solution

It has been believed that exposure to hypertonic solution during freezing is the major cause of the post-thaw cellular injury after freezing at low cooling rates. To develop a fundamental understanding of cell damage due to the hypertonic solution, we have developed a new perfusion technique that isolates the effect of exposure to hypertonic solutions from other effects associated with ice crystals. Using human prostatic adenocarcinoma cells, we characterized the post-hypertonic survival as a function of hypertonic solute concentration, the time of exposure, and the rates of increase and decrease in concentration. Also measured were the membrane permeability and the osmotically inactive volume of cells that are required for the description of osmotic change in cell volume.