Kinetics of Osmotic Water Movement in Chondrocytes Isolated from Articular Cartilage and Applications to Cryopreservation

The ability of chondrocytes to survive conditions encountered during cryopreservation depends on the responses of the cells to the physiochemical changes that result when water is removed from the environment of the cells in the form of ice. Cellular responses are therefore closely related to the osmotic permeability properties of the plasma membrane. In order to optimize the conditions for cryopreservation of chondrocytes, osmotic properties of the chondrocyte membrane were determined from the kinetics of volume change in hypertonic solutions at different temperatures. The hydraulic conductivity of the plasma membrane was 0.305 +/- 0.025 micron3/micron2/min/atmosphere at 24 degrees C, with an Arrhenius activation energy of 8.06 kcal/mol. These values are similar to those reported for other cell types, but the osmotically inactive volume of the chondrocytes (0.41 +/- 0.04) was significantly higher than for other cells, implying that chondrocytes have a higher dry weight ratio or that they contain a higher proportion of osmotically inactive or bound water. These results were used to calculate the osmotic responses of chondrocytes at low temperatures and to predict that the least damaging cooling rate for isolated chondrocytes in the absence of cryoprotective compounds is 10 degrees C/min. The ultimate goal of this study is the development of an analytical model applicable to the optimization of techniques for cryopreservation of intact cartilage and other tissues.