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为了探索冷冻对正常细胞和肿瘤细胞破坏的临界温度,本实验用三种正常细胞(乳猪肾、乳兔肾、兔睾丸)和宫颈癌细胞(HeLa细胞)作体外单层细胞实验。各种细胞培养成单层,经液氮冷冻后,加入~3H胸腺嘧啶核苷进行细胞DNA合成标记,测定DNA脉冲数作为生活细胞指标。另一组用S_(180)小白鼠肉瘤细胞悬液,经冷冻后注入动物体内,观察长瘤情况。实验结果表明,冷冻灭活正常细胞和肿瘤细胞的临界温度为-40℃维持3分钟,单层细胞仅需冻融一次,细胞悬液则需冻融三次。
In order to explore the critical temperature of freezing for normal cells and tumor cells, three normal cells (sugar kidney kidney, rabbit kidney, rabbit testis) and cervical cancer cells (HeLa cells) were used for in vitro monolayer cell experiments. All kinds of cells were cultured as monolayers, frozen in liquid nitrogen, and ~3H thymidine was added for cell DNA synthesis labeling, and the number of DNA pulses was measured as an indicator of living cells. The other group was treated with S_(180) mouse sarcoma cell suspension, frozen and injected into animals to observe the growth of the tumor. The experimental results showed that the critical temperature of frozen inactivated normal cells and tumor cells was maintained at -40°C for 3 minutes, and that the monolayer cells only required freezing and thawing. The cell suspension was frozen and thawed three times.