Factors influencing survival of mammalian cells exposed to hypothermia: II. Effects of various hypertonic media

Abstract

Survival of Chinese hamster lung (V79) cells, exposed as a function of time to hypothermia in tissue culture, in isosmotic and various hypertonic media was measured using a colony assay. The mechanism of hypothermie cell killing is different above and below 7 °C in this cell line. Addition of NaCl or mannitol to increase the tonicity to 400 mOsm greatly decreased the survival at 10 °C while addition of KCl had no significant effect. When these experiments were repeated at 5 °C, addition of either NaCl, KCl, or mannitol was detrimental to long-term cell survival. Furthermore, addition of mannitol to the medium did not improve survival when cells were stored at 7 °C. Addition of KC1 at 5 or 10 °C or NaCl at 5 °C only affected the cells' ability to accumulate sublethal damage, while addition of mannitol at 5 or 10 °C affected both of the above and the cold sensitivity of the cells. Addition of NaCl at 10 °C only affected the latter. These experiments suggest that prevention of cell swelling by these conditions, while possibly necessary during clinical hypothermie organ storage, is detrimental to single cell survival at these temperatures.