摘要
目的选择高效、稳定的细胞因子诱导杀伤细胞(CIK细胞)培养基及杀伤肝癌细胞的效靶比。方法采用RPMI1640、GT-T551两种培养基对5例健康人外周血单个核细胞进行体外常规诱导培养,两种培养基中分别加入0、200、500 IU/mL的IL-2,采用光学显微镜观察细胞形态变化,台盼蓝染色计数并绘制生长曲线,计算细胞扩增倍数。将CIK与肝癌7721细胞共培养,调整效靶比为10∶1、20∶1、40∶1、80∶1,72 h后进行CCK8检测,酶标仪测OD450值,计算杀伤率。结果与1640培养基比较,GT-T551培养基组各浓度IL-2下细胞扩增倍数增加(P均<0.05)。与效靶比80∶1时比较,效靶比为10∶1、20∶1、40∶1时CIK的杀伤率低(P均<0.05)。结论 GT-T551培养基加500 IU/mL IL-2的培养体系更有利于CIK细胞的增殖,CIK细胞与7721细胞的效靶比为80∶1时杀伤效应更强。
Objective The efficient and stable cytokine induced killer cell(CIK)culture medium and the effective target ratio of killing hepatocellular carcinoma cells were selected.Methods The peripheral blood mononuclear cells(PBMCs)from 5 healthy people were cultured in vitro by using RPMI1640 and GT-T551,and IL-2(0,200,and 500 IU/mL)was added to the two media,respectively.The morphology of the cells was observed by the optical microscope,and Trypan blue staining was used to count and draw the growth curve,and we calculated the cell amplification multiple.CIK was cocultured with liver cancer 7721 cell line,and the effector-target ratio was adjusted to 10∶1,20∶1,40∶1,and 80∶1.After72 h,we did the CCK8 test and measured OD450 by enzyme labeling instrument to calculate the killing rate.Results Compared with 1640 medium,the cell amplification multiples in the GT-T551 medium group at various concentrations of IL-2 increased(all P<0.05).Compared with the effector-target ratio of 80:1,the killing rate of CIK was low when the effector-target ratio was 10∶1,20∶1 and 40:1(all P<0.05).Conclusions The culture system of GT-T551 medium with 500 IU/mL IL-2 is more conducive to the proliferation of CIK cells.The cytotoxic effect reaches the peak when the effector-target ratio of CIK cells and 7721 cells is 80:1.
作者
李美鹤
戴尔珣
孙丽
侯雪芹
张汝建
LI Meihe;DAI Erxun;SUN Li;HOU Xueqin;ZHANG Rujian(Taishan Medical University,Taian 271000,China)
出处
《山东医药》
CAS
2019年第3期14-17,共4页
Shandong Medical Journal
关键词
培养基
效靶比
细胞因子诱导杀伤细胞
细胞增殖能力
细胞杀伤效应
culture medium
effector-target ratio
cytokine-induced killer cells
cell proliferation ability
cytotoxic effect
