两种培养基的培养体系对CIK细胞增殖和功能的影响

发布时间：2018-05-17 00:17

本文选题：白血病 + 幼红细胞　；参考：《重庆医学》2017年02期

【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。
[Abstract]:Aim to observe the proliferation of CIK cells induced by cytokines, and to detect the surface molecular phenotype of CIK cells and the cytotoxicity of K562 cells to K562 cells in vitro. Methods the mononuclear cells isolated from umbilical cord blood and autologous peripheral blood were cultured in H3 medium, and CIK cells were induced by adding recombinant human interferon- 纬 (IFN- 纬) and CD3 monoclonal antibody in vitro. On the 7th day, H3 and T551 were added to culture medium for 14 days. The proliferation ability of CIK cells was observed and the expression of CD3 + CD56 on the cell surface was detected by flow cytometry. The killing effect of two groups of culture medium on leukemia cell K562 was detected by CCK8 method. Results the dynamic counting and phenotypic analysis showed that the amplification times of CIK cells cultured in H3 and H3 ~ T551 (including total CIK cells in cord blood and total CIK cells in autologous CIK) reached 76.9 times and 62.3 times respectively on the 14th day. The contents of CD3 ~ CD56 ~ double positive cells in CIK cells of umbilical cord blood were 16.70 卤2.72% and 10.80 卤2.59%, respectively. The contents of CD3 ~ CD56 ~ double positive cells in autologous CIK cells were 11.23 卤6.64% and 10.70 卤6.42%, respectively. The killing rate of cord blood CIK cells cultured with H3 was 33.50 卤9.99g / L at 5:1, which was significantly higher than that of CIK cells cultured with H3T551 (20.3 卤6.76C). The difference was statistically significant (P < 0.05). The cytotoxicity of autologous CIK cells cultured with H3 and H3T551 was 59.67 卤27.59%, respectively. The difference was not statistically significant (P 0.080). Conclusion H3 cultured umbilical cord blood and autologous CIK cells have strong anti-leukemia cancer cell activity in vitro and can be used in clinical adoptive immunotherapy of leukemia.
【作者单位】：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;武汉汉密顿生物科技股份有限公司研发部;武汉大学第一临床学院肿瘤中心;武汉大学基础医学院;
【分类号】：R730.51

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