中文摘要：

在细胞的低温保存中,低温保护剂的种类与浓度对复温后的存活率有着重要影响。本文以人肝癌细胞Hep-G2为研究对象,利用慢速冷冻法,筛选最佳的冻存液配方。通过配比不同浓度的甘油、Me2SO以及在Me2SO中添加一定浓度的蔗糖、海藻糖,一周后复温细胞,对台盼蓝染色存活率、MTT存活率以及24 h贴壁率三种检测结果进行比较分析,结果表明：以Me2SO作为低温保护剂时,冻存液浓度为10%（v/v）的Me2SO复温后细胞的三种检测指标最优;以甘油作为低温保护剂时,冻存液浓度为20%（v/v）的甘油复温后细胞的三种检测指标最优;再将以上分别得到的最佳浓度（即体积浓度20%甘油、10%Me2SO）与5%Me2SO（v/v）＋0.3 mol/L蔗糖、5%Me2SO（v/v）＋0.3 mol/L海藻糖这四种低温保护剂进行冻存与比较,5%Me2SO（v/v）＋0.3mol/L海藻糖检测指标高于其他实验组,并且差异显著（P〈0.05）。最终得到5%Me2SO（v/v）＋0.3 mol/L海藻糖为慢速冷冻保存Hep-G2细胞的最优保护剂配方。

英文摘要：

It is well known that the type and concentration of cryoprotectant（ CPAs） exerts a significant influence on the survival rate of cells following cryopreservation. The optimal cryoprotectant for human hepatoma Hep-G2 cell with slow cooling method was determined in the study. The Cells were frozen in Me2 SO,glycerol,sucrose and trehalose at different concentration and combination,then stored in a- 80 ℃ freezer for one week. The survival rate was assessed by trypan blue dye exclusion test,MTT assay and 24 h attachment assay.The results suggested that 10%（ v / v） Me2 SO provide effective protection among Me2 SO groups; 20%（ v / v） glycerol provide effective protection among glycerol groups; when sugars were added,5% Me2SO（ v / v） ＋ 0. 3 mol / L trehalose provide effective protection than the other groups（ P 〈0. 05）. In conclusion,5% Me2SO（ v / v） ＋ 0. 3 mol / L trehalose was the presumptive optimal cryoprotectant for human hepatoma Hep-G2 cell during slow freezing.