人肝癌细胞Hep-G2的冷冻干燥保存初探

冷冻干燥是长期有效保存细胞的手段之一。本文以肝癌细胞Hep-G2为研究对象,选取了不同体积分数的Me2SO、丙三醇及不同质量浓度的PVP、复方保护剂进行冷冻干燥保存,筛选出最佳保护剂配方。此外,将细胞在不同摩尔浓度的海藻糖溶液中孵育,通过检测细胞回收率、存活率和24 h贴壁率,探究胞内海藻糖对细胞冷冻干燥的影响。结果表明:添加40%PVP(w/v)+10%甘油(v/v)+15%FBS(v/v)+20%海藻糖(w/v)保护剂的细胞,在复水后回收率、存活率和24 h贴壁率分别为29.58%、42.18%和18.71%,与对照组相比三项指标均得到有效提高,该种保护剂的效果最佳;当胞外海藻糖摩尔浓度为800 mmol/L时,冻干效果最好,肝癌细胞Hep-G2在复水后回收率、存活率和24 h贴壁率分别为27.81%,66.65%和33.68%,存活率和贴壁率显著高于其他组。

Preliminary Research on Freeze-drying Preservation of Human Hepatoma Hep-G2 Cells

Freeze-drying is a long-term and effective method for cell preservation. The lyoprotectant can have a significant effect on cell freeze-drying. In this study, different volume concentrations of Me2SO, glycerol, and different mass concentrations polyvinylpyrrolidone (PVP), compound lyoprotectant for lyophilizing human Hepatoma Hep-G2 cells were considered. The effect of intracellular trehalose on the lyophilized cells was investigated by determining the cell-recovery rate, survival rate, and 24 h attachment rate. The results show that, compared with the control group, the cells added in a 40% PVP (w/v) + 10% (v/v) glycerol + 15% fetal bovine serum (FBS) (v/v) + 20% trehalose (w/v) solution received better protection. The cell recovery rate, survival rate, and 24 h adherent rate are 29.58%, 42.18%, and 18.71%, respectively. When the extracellular trehalose molar concentration is 800 mmol/L, the Hepatoma Hep-G2 cell-recovery rate, survival rate, and 24 h adherent rate are 27.81%, 66.65%, and 33.68%, respectively. It is significantly better than the control group and has the best protection for cell freeze-drying.