Study on Cryopreservation of Human Hepatoma Hep-G2 Cell

Yao Lan; Liang Wei; Liu Baolin

doi:10.3969/j.issn.0253-4339.2015.02.095

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Study on Cryopreservation of Human Hepatoma Hep-G2 Cell

更新时间：2025-09-03

- Study on Cryopreservation of Human Hepatoma Hep-G2 Cell
- Journal of Refrigeration Vol. 36, Issue 2, Pages: 95-100(2015)
- 作者机构：
  
  上海理工大学生物系统热科学研究所
- 作者简介：
- 基金信息：
- DOI：10.3969/j.issn.0253-4339.2015.02.095
  CLC：
- Published：2015
- 稿件说明：
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Yao Lan, Liang Wei, Liu Baolin. Study on Cryopreservation of Human Hepatoma Hep-G2 Cell[J]. Journal of Refrigeration, 2015, 36(2): 95-100.
DOI：

Yao Lan, Liang Wei, Liu Baolin. Study on Cryopreservation of Human Hepatoma Hep-G2 Cell[J]. Journal of Refrigeration, 2015, 36(2): 95-100. DOI： 10.3969/j.issn.0253-4339.2015.02.095.

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。

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.

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Related Institution

上海理工大学

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Institute of Biothermal and Technology, University of Shanghai for Science and Technology

Institute of Biothermal Science and Technology, University of Shanghai for Science and Technology

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