两种培养基的培养体系对CIK细胞增殖和功能的影响  

Effects of two different cell culture medium on proliferative and functional activities of cytokine-induced killer cells

作　　者：宋文玲[1] 韩蓉[1] 张曼[1] 张权 姚惟琦[2,3] 武栋成[2,4] 

机构地区：[1]湖北省武汉市黄陂区人民医院暨江汉大学附属第三医院肿瘤三科,430060 [2]武汉汉密顿生物科技股份有限公司研发部,武汉430075 [3]武汉大学第一临床学院肿瘤中心,武汉430060 [4]武汉大学基础医学院,武汉430072

出　　处：《重庆医学》2017年第2期215-217,222,共4页Chongqing medicine

摘　　要：目的观察细胞因子诱导的杀伤细胞(CIK细胞)增殖情况,检测CIK细胞的表面分子表型和体外对白血病细胞K562的杀伤作用。方法通过用H3培养基培养的分别来源于脐带血和患者自体外周血分离的单个核细胞(PBMC),添加重组人γ干扰素(IFN-γ)与CD3单抗,体外诱导CIK细胞,在培养的第7天分别加入H3培养基和T551培养基继续诱导培养至14d。计数观察CIK细胞增殖能力,流式检测细胞表面CD3、CD56的表达情况,CCK8法检测两组培养基条件下对白血病细胞K562的杀伤效果。结果动态计数及表型分析结果表明,H3及H3^+T551培养的CIK细胞扩增倍数(包括脐带血CIK总细胞数和自体CIK总细胞数)在第14天均分别达到76.9倍、62.3倍;两种培养条件下脐带血CIK细胞中CD3^+CD56^+双阳性细胞含量分别是(16.70±2.72)%和(10.80±2.59)%,自体CIK细胞中CD3^+CD56^+双阳性细胞含量分别是(11.23±6.64)%和(10.70±6.42)%;体外杀瘤实验表明,当效靶比为5∶1时,H3培养的脐带血CIK细胞杀伤率达到(33.50±9.99)%,显著高于H3^+T551培养脐带血CIK细胞的(20.3±6.76)%,差异有统计学意义(P=0.011),而H3和H3^+T551培养的自体CIK细胞杀伤率分别是(59.67±27.59)%和(42.13±19.47)%,差异无统计学意义(P=0.080)。结论 H3培养的脐带血和自体CIK细胞具有较强的体外抗白血病癌细胞活性,可应用于临床上白血病的过继性免疫治疗。Objective To observe the proliferation ability of cytokine-induced killer（CIK）cells and to detect surface molecule phenotype of CIK cells and in vitro killing effect to leukemia K562 cells.Methods The cord blood lymphocytes and peripheral blood mononuclear cells（PBMCs）from patients were cultured by the H3 medium and added with human recombinant interferon（IFN）-γand CD3 monoclonal antibodies（mAb）for inducing CIK cells in vitro.The H3 medium and T551 medium were respectively added on 7dof culture.The induction and culture were continued until 14 d.The CIK cells proliferation ability was observed by cell count and the expressions of CD3 and CD56were detected by flow cytometry.The killing effect of CIK cells on leukemia cells was tested by CCK8 assays.Results The dynamic counting and phenotype analysis results revealed that the CIK cells amplification times（including cord blood CIK total cells and autologous CIK total cells）cultured by H3 and H3＋T551reached 76.9times and62.3times on 14drespectively;the CD3＋CD56＋double positive cells contents of cord blood CIK cells under the two kinds of culture condition were（16.7±2.7）% and（10.8±2.6）% respectively,while which of autologous CIK cells were（11.2±6.6）% and（10.7±6.4）% respectively;the in vitro killing-tumor test revealed that when the effector to-target ratio was 5：1,the cell cytotoxic activity of cord blood CIK cells cultured by H3reached（33.50±9.99）%,which was significantly higher than（20.3±6.76）% of cord CIK cells cultured by H3＋T551（P=0.011）,while which of autologous CIK cells cultured by H3＋T551 were（59.67±27.59）% and（42.13±19.47）% respectively,but the difference was not significant（P=0.080）.Conclusion Cord blood and autologous CIK cells incubated by H3 have stronger in vitro anti-leukemia cells activity and can be used in the adoptive immunotherapy of leukemia in clinic.

关 键 词：白血病 幼红细胞 急性 CIK细胞 白血病细胞K562 H3培养基 T551培养基 杀伤率 

分 类 号：R730.51[医药卫生—肿瘤]