三种试剂用于两种肿瘤细胞活性检测的比较研究

Comparison of three reagents for detecting two kinds of tumor cell activity

目的通过对三种试剂用于两种肿瘤细胞活性检测性能的综合对比,提供有理论依据的实践参考。方法分别以贴壁细胞Hela和悬浮细胞SU-DHL-2为研究对象,对CCK-8、alamarBlue和CellTiter-Glo三种常用试剂的最佳条件进行探索。结果对于两种细胞,三种试剂在一定条件下均有较好的线性区间,Pearson相关系数r>0.99;CCK-8用于检测Hela的细胞数目为8 000个/孔,用于SU-DHL-2为28 000个/孔,最佳孵育时间为1h;alamarBlue可检测两种细胞的数目分别为14 000个/孔和16 000个/孔,最佳孵育时间分别为6h和2h;CellTiter-Glo可检测的细胞数目均为50 000个/孔,孵育时间为10min。结论 CCK-8和alamarBlue成本较低,孵育时间较长,线性范围较窄,实验前需要耗费时间摸索最佳条件;CellTiter-Glo线性范围较宽,检测效率和准确性最高,但是需要专门的检测设备,成本较高。

Objective To provide a comprehensive comparison and optimized protocol of three different cell activity detection reagents for two kinds of tumor cell.Methods The optimal conditions for the three common reagents of CCK-8,alamarBlue and CellTiter-Glo were explored respectively with adherent cell Hela and suspension cell SU-DHL-2 as research subjects.Results For both kinds of cells,the three reagents had a good linear range under certain conditions,and all the Pearson correlation coefficient r>0.99.For CCK-8,the optimal alternative used to detect the Hela and SU-DHL-2 cell number were 8 000 cells per well and 28 000 cells per well respectively,and the best incubation time was 1h.For alamarBlue,the optimal alternative used to detect the number of two kinds of cells were 14 000 cells per well and 16 000 cells per well,and the optimal incubation time were 6h and 2h respectively.For CellTiter-Glo,up to 50 000 cells per well and 10 minutes incubation time had been detected.Conclusion CCK-8 and alamarBlue have lower cost,longer incubation time and narrower linear range,but the optimized protocol should be taken by pre-experiment with long time spending;CellTiter-Glo has a wide linear range,and the highest detection efficiency and accuracy,but the cost is higher because it requires special detection equipment.