Regulatory volume decrease in COS-7 cells at 22 °C and its influence on the Boyle van’t Hoff relation and the determination of the osmotically inactive volume

摘要

Cryobiological analyses assume that the direction and rate of water movements across cell membranes and equilibrium cell volumes are determined solely by differences in the chemical potentials of intra- and extracellular water. A consequence of this assumption is that cells obey the Boyle van’t Hoff (BvH) law which states that cell volumes are a linear function of reciprocal osmolality. Extrapolation of the BvH plot to infinite osmolality yields a quantity b, the fractional volume of the cell occupied by solids. In many cells, however, a cell volume excursion above the isotonic volume initiates an energy-requiring response that causes the swollen cells to shrink back to or towards isotonic volume. It is referred to as Regulatory Volume Decrease (RVD). We have observed a strong RVD in COS-7 cells. If not eliminated by keeping exposure times short, this RVD produces a b that is 60% too high (0.48 vs. 0.30) These results indicate the importance of examining cells for volume regulatory mechanisms before performing measurements to determine their osmotic parameters.