【摘要】 MTT检测的OD值与细胞量之间呈显著的线性正相关(R2>0.96),能较好地反映活细胞密度。MTT的作用浓度及反应时间对此OD值均有较大影响,根据实验结果在细胞的MTT检测中,采用0.5 mg/mL为MTT作用浓度,4 h为反应时间。细胞接种量对最终的细胞活力有一定影响,根据实验结果在鱼类细胞毒性实验中,选取每孔2.0×104为最初接种量(初始密度为105cell/mL),对照SMMC7721细胞则选择每孔0.5×104细胞接种量(初始密度为2.5×104cell/mL)。通过优化的MTT比色法检测抗生素和多环芳香烃等药物对细胞的毒性,药物的剂量-效应曲线以Logistic模型拟合,获得实验药物对不同细胞的半致死浓度IC50,结果表明,多环芳香烃的毒性通常大于抗生素,研究还发现药物对C IK、GCL、PCK细胞的IC50显著高于SMMC7721细胞(P<0.05)。更多还原

【Abstract】 OD value in MTT assay had remarkable linear correlation with cell number,so it could well present the density of viable cells.Concentration and response time of MTT had great influence on that value,and the best of them were chosen to be 0.5 mg/mL and 4 h respectively according to the result in MTT test.Inoculating cell number had certain effect on the viability of cells.It was optimum that fish cell lines inoculated by 2.0×104 cells in each well(equal to 105 cell/mL) and the control SMMC7721 inoculated by 0.5×104 cells(equal to 2.5×104 cell/mL).Examined with the optimized MTT method and fit by the Logistic model,IC50 of various drugs to inhibit cell viability were acquired.IC50 values revealed that polyaromatic hydrocarbons had higher cytotoxicity than antibiotics.Furthermore,IC50 values of drugs in CIK,GCL and PCK cell lines were higher than those in SMMC7721(P<0.05).更多还原