A method for maintaining constant ethanol concentrations in cell culture media.

The present study reports on the development of a model for maintaining constant ethanol concentrations over time in cell culture media. When neuroblastoma x glioma cells (NG 108-15) were grown in ethanol containing media under standard cultivation conditions in the incubator at 37 degrees C, a 90% evaporation was observed after 24 hr. To counteract evaporation, the cell culture dishes were placed inside polystyrene boxes together with an open dish containing an appropriate amount of ethanol. By using such procedure, the decrease in ethanol concentration in the culture media was completely avoided. Cultivating cells in ethanol-free media inside sealed plastic boxes did not change their viability, growth rate, protein and phospholipid composition of the cells or the pH of the media, compared to cultures grown outside the boxes.