神经癌细胞PC12对二甲基亚砜耐受剂量的分析

Dose Analysis of Dimethyl Sulfoxide to Morphology and Growth of PC12 Cell

通过观察不同剂量二甲基亚砜(dimethyl sulfoxide DMSO)作用下PC12细胞形态和生长状态的变化,确定PC12细胞对DMSO的耐受剂量.按照一定浓度梯度(V/V)将DMSO加入培养液干预后的24、48和72 h于倒置相差显微镜下观察AO/EB染色前后细胞的形态特征,同时用四甲基偶氮唑蓝(MTT)显色法检测细胞的生存率和计算半数抑制浓度(50%inhibiting concentration,IC50).结果表明:DMSO浓度为14 mL/L时,24 h内90%的PC12细胞生长正常,形态完整,活力与对照组比较无差别,浓度提高到20 mL/L时,24 h时细胞生存率可下降27%,随着作用时间延长,细胞生存率进一步降低;24、48和72 h对应的IC50值分别为31.4、23.9和19.2 mL/L.因此,对PC12细胞而言,24 h内的应用终浓度应低于14 mL/L,干预时间如需延长,DMSO的应用剂量应相应降低.

The morphologic changes and growth status of PC 12 cell were observed after being intervened by different concentrations of Dimethyl Sulfoxide and the safe concentrations were ascertained. AO/PI staining was chosen to show the morphologic changes of PC12 and cellular pictures were taken before and after being stained when cells were cultured full of 24, 48 and 72 h respectively. At the same time, cell viability and ICs0 were quantitated based on the assay of MTT. Except for regular growth, 90% of the PC12 also could keep perfect structure and the same cell viability as the control in 24 h if the concentration of DMSO was 14 mL/L. If the concentration of DMSO was up to 20 mL/L, the cell viability would be down to 73% in 24 h. The longer the time, the more decreased the percentages of cell viability. The IC50 values corresponding to 24,48 and 72 h were 31.4, 23.9 and 19.2 mL/L respectively. The safe concentration of DMSO to PC12 was not above 14 mL/L in 24 hours, if needing extension of intervention time, the corresponding concentrations of DMSO should be reduced.