人肝癌细胞Hep-G2的冷冻干燥保存初探
Preliminary Research on Freeze-drying Preservation of Human Hepatoma Hep-G2 Cells

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。
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