Influence of Hypothermia on Cell Volume and Cytotoxic Swelling of Glial Cells in Vitro

Abstract

In view of the increasing significance of mild hypothermia (32 °C) as an efficient procedure of neuroprotection, the present study was performed to examine the influence of this level of hypothermia on the volume of glial cells under physiological as well as under pathological conditions.

The influence of mild (32 °C) and moderate (27 °C) hypothermia on cell volume and cell viability of C6 glioma cells was studied for 60 minutes in vitro. Cells were suspended in an incubation chamber under continuous control of temperature, pH and pO₂. Cell volume was measured by an advanced Coulter system. Hypothermia itself was causing significant cell swelling in a dose-dependent manner, which could be prevented by omission of Na⁺-ions from the suspension medium, while the replacement of Cl^--ions failed to prevent cell swelling from hypothermia. Inhibition of the Na⁺/H⁺- antiporter with EIPA (5N-ethyl-n-isopropyl-amiloride, 50 µM) was significantly reducing the hypothermia induced cell swelling, indicating activation of the Na⁺/H⁺-antiporter.

Conversely, mild or moderate hypothermia failed to prevent cell swelling from lactic acid, arachidonic acid or glutamate, i.e. agents which are mediating the development of cytotoxic brain edema in vivo in cerebral trauma, ischemia and other acute insults. The findings indicate that cerebral protection by hypothermia in vivo is most likely not attributable to an inhibition of cytotoxic brain edema. Further investigations, however, are required in vivo and in vitro to elucidate the hypothermia-induced swelling of glial cells in more detail, e.g. as to the role of the Na⁺/H⁺-antiporter.