不同培养基中CIK的生物学特性及其对肿瘤细胞的杀伤效果

Research on Biological Characteristics and Cytotoxicity of CIK Cells in Different Culture Systems

目的比较健康人和肿瘤患者细胞因子诱导的杀伤细胞(cytokine-induced killer,CIK)在淋巴细胞无血清培养基及普通无血清培养基中的细胞形态、体外扩增速度、细胞表型及其对肿瘤细胞株的杀伤活性。方法取健康人和肿瘤患者外周血各3例,经密度梯度离心获得外周血单个核细胞(PBMC),分别在淋巴细胞无血清培养基及普通无血清培养基中,加入基因重组人干扰素γ(rhIFN-γ)、抗人CD3单克隆抗体(OKT3)和基因重组人白细胞介素2(rhIL-2),体外诱导CIK细胞,比较细胞的形态、扩增速度、细胞表型及杀伤活性。结果在不同培养基中健康人外周血CIK细胞的扩增速度、流式细胞学检测的CD3+CD8+和CD3+CD56+细胞百分比以及对K562和HL-60肿瘤细胞株的杀伤活性均显著高于肿瘤患者,差异有统计学意义(P<0.05);两种来源的PBMC在不同的培养基中经过诱导获得的CIK细胞在增殖速度、流式细胞学分析结果及杀伤活性方面差异亦有统计学意义(P<0.05)。结论在不同的培养基中肿瘤患者的PBMC经诱导培养获得的CIK细胞可有显著差别,为临床进一步提高CIK细胞的治疗效果提供了实验依据。

Objective To observe the changes in morphology,phenotype,proliferation activity and cytotoxicity of cytokine induced killer（CIK） cells which were cultured in lymphocytic serum-free medium or in general purpose serum-free medium and from healthy people or tumor patients,and then to investigate the efficiency of CIK cells in cancer immunotherapy.Methods Peripheral blood mononuclear cells（PBMC） were isolated by density gradient centrifugation derived from three healthy subjects and three tumor patients,CIK cells were generated from PBMC in lymphocytic serum-free medium A or B and were incubated in vitro in the presence of IFN-γ followed by OKT3 and IL-2.Proliferation,Morphology,phenotypes,and the cytotoxicity activity were compared.Results The proliferation speed,the percentage of CD3^＋CD8^＋ and CD3^＋CD56^＋ cells,as well as the cytotoxicity activity of CIK cells originated from peripheral blood of healthy people were significantly higher than those of tumor patients（P〈0.05） under different culture systems;CIK cells generated from different sources of PBMC also showed significant differences at proliferation speed,phenotypes and cytotoxicity activity when cultured under different systems.Conclusion CIK cells generated from PBMC of tumor patients have significant differences under different culture systems,and provide further experimental basis for improving the clinical curative effect of CIK cells.