| 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24h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)+0.3 mol/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。 |
| 关键词: 低温保存 低温保护剂 糖类 存活率 |
| DOI： |
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| 基金项目:国家自然科学基金（51076108）资助项目。 |
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| Study on Cryopreservation of Human Hepatoma Hep-G2 Cell |
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Yao Lan, Liang Wei, Liu Baolin
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| (Institute of Biothermal and Technology，University of Shanghai for Science and Technology) |
| Abstract: |
| 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 Me2SO, 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) Me2SO provide effective protection among Me2SO groups; 20% (v/v) glycerol provide effective protection among glycerol groups; when sugars were added, 5%Me2SO(v/v)+0.3mol/L trehalose provide effective protection than the other groups（P<0.05）. In conclusion, 5% Me2SO (v/v)+0.3mol/L trehalose was the presumptive optimal cryoprotectant for human hepatoma Hep-G2 cell during slow freezing. |
| Key words: cryopreservation cryoprotectants sugar viability |
