人肝癌细胞Hep-G2的冷冻干燥保存初探

发布时间：2018-11-27 14:23

【摘要】：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。
[Abstract]:Freeze-drying is one of the means of long-term preservation of cells. In this paper, we select different volume fractions of Me2SO, glycerol and different mass concentrations of PVP and compound protectants for freeze-drying and storage, and screen out the optimal formulation of protective agent. In addition, the cells were incubated in a different molar concentration of trehalose solution, and the effect of intracellular trehalose on the freeze-drying of the cells was explored by the detection of cell recovery, survival and 24 h adherence. The results showed that 40% PVP (w/ v) + 10% glycerol (v/ v) + 15% FBS (v/ v) + 20% trehalose (w/ v) protectants were added. The recovery rate, survival rate and the adherence rate of 24 h were 29. 58%, 42. 18% and 18. 71%, respectively. When the molar concentration of the extracellular trehalose is 800 mmol/ L, the freeze-drying effect is the best, and the recovery rate, the survival rate and the 24-hour adherence rate of the Hep-G2 in the liver cancer cell are 27. 81%, 66. 65% and 33. 68%, respectively, and the survival rate and the adherence rate are significantly higher than the other groups.
【作者单位】： 上海理工大学医疗器械与食品学院;
【基金】：国家自然科学基金(51376132)资助项目~~
【分类号】：R73-3
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本文编号：2361106