8种增溶剂/溶剂对中国仓鼠卵巢(CHO)细胞活性影响的观察

Influence of 8 Solubilizers/Solvents on the Viability of Chinese Hamster Ovary(CHO) Cells

目的通过观察8种增溶剂/溶剂对中国仓鼠卵巢(CHO)细胞活性的影响,推断其用于CHO细胞染色体畸变试验时的适用浓度。方法以增溶剂/溶剂作用3 h(+S9,-S9)和24 h(-S9)对CHO细胞进行染毒;刃天青法测定染毒细胞活性;Logit法计算半数抑制浓度IC50和5%抑制浓度IC5。结果各增溶剂/溶剂在作用3 h(+S9,-S9)和24 h(-S9)后,对CHO细胞的IC50和IC5影响各异,其中非离子型增溶剂吐温-20的IC50小于0.5 mg/mL(24 h作用组<0.05 mg/mL);-S9条件下3 h和24 h的IC5分别为1.0和0.1(丙酮)、1.1和0.3(DMSO)、0.1和<0.01(吐温-20)、0.1和<0.01(吐温-60)、1.1和0.4(甲醇)、0.5和0.6(95%乙醇)、0.5和0.3(乙酸乙酯)和0.5和1.5 mg/mL(丙三醇)。结论 8种增溶剂/溶剂能够不同程度影响CHO细胞的活性,日常检测工作中可参考其IC50和IC5以确定使用剂量。

Objectives To analyse the effects of 8 solubilizers/solvents on the viability of Chinese Hamster Ovary（CHO） cells to deduce the applicable doses in CHO chromosomal aberration test.Methods CHO cells were treated with 8 solubilizers/solvents for 3 h（＋S9-S9） and 24 h（-S9） respectively.The viability of CHO was detected by resazurin method.Logit method was used to calculate the 50% inhibitory concentration（IC50） and 5%inhibitory concentration（IC5）.Results The IC50 and IC5 values of 8 solubilizers/solvents to CHO cell（treated with or without S9 mixture for 3 h,and treated with S9 mixture for 24 h） were variable.The IC50 values of non-ionic solubilizer Tween-20 were less than 0.5 mg/mL（especially less than 0.05 mg/mL in the 24 h treatedt group）.IC5 values in 3 and 24 h treated groups（without S9 mixture） were 1.0 and 0.1（acetone）,1.1 and 0.3（DMSO）,0.1 and less than 0.01（Tween-20）,0.1 and〈0.01（Tween-60）,1.1 and 0.4（methanol）,0.5 and 0.6（95% ethanol）,0.5 and 0.3（ethyl acetate）,and 0.5 and 1.5 mg/mL（glycerol） respectively.Conclusions There were some influences of 8 solubilizers/solvents on the viability of CHO in different degrees.The dosage used in the usual examination may refer to the IC50 and IC5 of each solubilizer/solvent.