玻璃化低温保存卵巢癌细胞的配方及优化降温过程参数

Formulation for vitrification solution and cooling process parameters in cryopreservation of human ovarian cancer COC1 cells

实验采用玻璃化低温保存方法和程序降温方法来研究COC1细胞的低温保存效果。玻璃化低温保存:采用3种玻璃化溶液,①质量浓度为400g/L聚乙烯基吡咯烷酮+质量浓度为200g/L蔗糖+质量浓度为100g/L甘露醇。②VS55。③质量浓度为300g/L聚乙烯基吡咯烷酮+质量浓度为200g/L海藻糖。每种溶液以不同的比例与细胞悬液混合均匀,投入液氮保存。程序降温法:以甘油和二甲基亚砜为低温保护剂,分别以不同的比例添加到含有细胞悬液的冻存管中。然后采用两步降温法,投入液氮保存。结果玻璃化溶液(质量浓度为300g/L聚乙烯基吡咯烷酮+质量浓度为200g/L海藻糖)保存COC1细胞可以获得最高细胞存活率,且玻璃化方法取得的存活率高于程序降温法。初步表明,用玻璃化方法长期保存卵巢癌细胞具有一定的可行性和前景,使COC1细胞能够最大限度的保持较好的生理功能。

This experiment was to study the cryopreservative effect of vitrification and programmed cooling method on human ovarian cancer COC1 cells. Three kinds of vitrification solutions were used in vitrification. （1）400 g/L polyvinylpyrrolidone ＋ 200 g/L sucrose ＋ 100 g/L mannitol; （2）VS55; （3）300 g/L polyvinylpyrrolidone ＋ 200 g/L trehalose. The three kinds of vitrification solutions were mixed with COC 1 cells suspensions at different proportion, respectively. Then the samples were preservd in liquid nitrogen. Programmed cooling method： The glycerol and Dimethylsulfoxide, as cryoprotective agents, were mixed with COC1 cells suspensions at different concentration, respectively. After cooled by two step method, the samples were preserved in liquid nitrogen. The experiment found that the vitrification solution composed of 300 g/L polyvinylpyrrolidone ＋ 200 g/L trehalose could obtain the highest cell viability of human ovarian cancer COC 1 cells. And the viability of COC 1 cells preserved by vitrification was higher than that preserved by programmed cooling method. The results indicted that long-term cryopreservation of human ovarian cancer COC 1 cells by using vitrification has the feasiblility and good prospects, and the vitrification can maintain the physiological function of COC 1 cells to the maximum extent.