Rapid decrease in cellular sodium and chloride content during cold incubation of cultured liver endothelial cells and hepatocytes

Hypothermia, as used for organ preservation in transplantation medicine, is generally supposed to lead to an intracellular accumulation of sodium, and subsequently of chloride, via inhibition of the Na+/K+-ATPase. However, on studying the cellular sodium concentration of cultured liver endothelial cells using fluorescence microscopy, we found a 55% decrease in the cellular sodium concentration after 30 min of cold incubation in University of Wisconsin (UW) solution. To confirm this surprising result, we set up a capillary electrophoresis method that allowed us to determine the cellular contents of inorganic cations and of inorganic anions. Using this method we measured a decrease in the cellular sodium content from 104+/-11 to 55+/-4 nmol/mg of protein, accompanied by a decrease in the chloride content from 71 +/- 9 to 25 +/- 5 nmol/mg of protein, after 30 min of cold incubation in UW solution. When the endothelial cells were incubated in cold Krebs-Henseleit buffer or in cold cell culture medium instead of UW solution, similar early decreases in cellular sodium and chloride contents were observed, thus excluding the possibility of the decreases being dependent on the preservation solution used. Furthermore, experiments with cultured rat hepatocytes yielded a similar decrease in sodium content during initiation of cold incubation in UW solution, so the decrease does not appear to be cell-specific either. These results suggest that, contrary to current opinion, sodium efflux predominates over sodium influx during the early phase of cold incubation of cells.