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：2024-07-18

- Study on Cryopreservation of Human Hepatoma Hep-G2 Cell
- Journal of Refrigeration Vol. 36, Issue 2, (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).
DOI：

Yao Lan, Liang Wei, Liu Baolin. Study on Cryopreservation of Human Hepatoma Hep-G2 Cell[J]. Journal of refrigeration, 2015, 36(2). 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 Author

胥义

何伟

占太杰

崔梦冬

刘林峰

Liu Baolin

Ye Ping

Liang Wei

Related Institution

上海理工大学

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

Institute of Biothermal Science and Technology, University of Shanghai for Science and Technology, Shanghai Technical Service Platform for Cryopreservation of Biological Resources, Shanghai Collaborative Innovation Center for Tumor Treatment with Energy

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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