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目的：研究冻融后癌细胞不同时间的生存率改变，建立准确测定冷冻对癌细胞损伤效应的方法。方法：采用体外培养的６种口腔癌细胞和两种正常细胞进行冷冻处理，ＭＴＴ法测定不同温度的生存率，选择－４０℃为敏感温度进行冻融后不同时间的冷冻后生存率测定。结果：不同温度冷冻癌细胞后测定结果显示：０℃～－３０℃细胞无明显损伤，－４０℃～－５０℃死亡率明显增加，达到４０％～６０％、－５０℃～－９０℃细胞死亡率未见明显增加。－４０℃冷冻后细胞死亡率随时间延长而有所增加，８ｈ达６０％～８５％，２４ｈ达８４％～９４％．结论：冷冻对癌细胞有明确杀伤作用，敏感温度在－４０℃～－５０℃。冻融后部分存活细胞的损伤无法修复，细胞死亡率随时间延长而缓慢增加，２４ｈ达到最大值。
OBJECTIVE: To study the changes of survival rate of cancer cells after freezing and thawing, and to establish an accurate method to determine the effects of freezing on cancer cells. Methods: Six kinds of oral cancer cells and two normal cells were cultured in vitro for freezing treatment. MTT method was used to measure the survival rate at different temperatures. The survival rate after frozen at -40 ℃ was determined. Results: The results of frozen-thawed cells at different temperatures showed that there was no obvious cell damage at 0 ℃ ~ -30 ℃, and the mortality rate at -40 ℃ ~ -50 ℃ increased significantly, reaching 40% ~ 60%, at -50 ℃ ~ -90 ℃ No significant increase in mortality. The cell death rate after freezing at -40 ° C increased with time, reaching 8% to 85% at 8 h and 84% to 94% at 24 h. Conclusion: Cryotherapy has a clear killing effect on cancer cells. The sensitivity temperature is between -40 ℃ and -50 ℃. After freezing and thawing, some of the surviving cells could not be repaired. The cell death rate increased slowly with time and reached the maximum at 24h.