CCK-8法与LDH法检测CIK细胞毒活性的比较

摘要

Objective:To investigate the optimal conditions of CCK-8 and LDH assay for detecting CIK cell cytotoxic activity.Methods:Peripheral blood mononuclear cells(PBMCs)were isolated by Ficoll-Paque gradient centrifugation and CIK cells were amplified by cytokine.After culturing 14 days,the cytotoxicity of CIK cells to prostate cancer cells was detected by CCK-8 and LDH assay.Results:The best detection time of CCK-8 method was 3~4 h after adding CCK-8 reagent,and the suitable cell number range was 0.32×103~2.0×104 cells/hole.Conclusion:CCK-8 is a better method than LDH in stability,accuracy and operation to detect the cytotoxic activity of immune cells.%目的:探讨CCK-8法和LDH法检测CIK细胞活性的最佳条件.方法:经密度梯度法分离得到人外周血中单个核细胞,经细胞因子刺激培养14 天收获CIK细胞,采用CCK-8法与LDH法检测CIK细胞对前列腺癌细胞毒活性.结果:CCK-8法的最佳检测时间为加入CCK-8试剂后3~4 h,适宜细胞数量范围为0.32×103~2.0×104个/孔.结论:CCK-8法较LDH法稳定性好,准确性好,操作简便,是一种优于LDH法的检测免疫细胞毒活性的方法.