恶性肿瘤患者自体CIK细胞的诱导培养及表型鉴定  

In vitro culture and phenotypic analysis of autologous cytokine induced-killer cells derived from patients with malignancy

作　　者：喻皇飞[1] 方宁[1] 章涛[1] 陈代雄[1] 范振海[1] 杨卫兵[2] 宫黎明[1] 刘金伟[1] 余丽梅[1] 

机构地区：[1]遵义医学院附属医院贵州省细胞工程重点实验室,贵州遵义563003 [2]遵义医学院附属医院肿瘤医院,贵州遵义563003

出　　处：《遵义医学院学报》2011年第4期354-356,360,共4页Journal of Zunyi Medical University

基　　金：贵州省科技厅平台建设项目资助209.001.106.01

摘　　要：目的观察恶性肿瘤患者外周血来源的细胞因子诱导杀伤(CIK)细胞体外培养形态及免疫表型变化。方法采用费森尤斯CEM.TEC血细胞分离机采集恶性肿瘤患者自体外周血单个核细胞,经rhIFN-、rhIL-2及CD3McAb联合诱导培养后,显微镜下观察诱导细胞形态,流式细胞术分析其免疫表型。结果连续诱导培养10 d后CIK细胞呈团样生长;至13 d时,CD3+、CD3+CD8+和CD3+CD56+细胞比例分别从诱导前的(44.8±11.3)%、(21.1±8.5)%和(3.4±10.7)%增加至(72.1±13.8)%、(51.1±17.9)%及(10.7±4.4)%(<0.05);而CD3+CD4+细胞比例为(19.2±8.0)%,明显低于诱导前(28.7±6.4)%(<0.05)。结论来自恶性肿瘤患者的自体外周血单个核细胞可成功诱导为富含CIK细胞的肿瘤杀伤细胞。Objective To observe the morphology and immunophenotypic properties of autologous cytokine induced killer（CIK） cells derived from the peripheral blood of patients with malignant tumor.Methods Peripheral blood mononuclear cells（PBMNCs） separation of patients with malignant tumor were carried out by Fresenius Kabi CEM.TEC equipment,then the PBMNCs were cultured and induced into CIK cells with the combination of cytokines including recombinant human interferon-γ（rhIFN-γ）,recombinant human interleukin-2（rhIL-2） and anti-CD3McAb.Results Before and after induction,the cells morphology was observed under microscope,and the immunophenotypic properties were examed by flow cytometry.CIK cells presented a cluster like growth manner after 10 days continuously induction.The percentage of CD3＋,CD3＋CD8 and CD3＋CD56＋ cells were increased significantly from（44.8±1.3）%,（21.1±.5）%,（3.4±0.7）% to（72.1±3.8）%,（51.1±7.9）%,（10.7±.4）%,respectively at 13 days post-induction（P0.05）.Whereas,the percentage of CD3＋CD4＋ cells were decreased from（28.7±.4）% to（19.2±.0）%（P0.05）.Conclusion Autologous PBMNCs derived from cancerpatients could be successfully induced into cancer killer cells that abundantly contained CIK cells.

关 键 词：肿瘤 细胞因子诱导杀伤细胞 免疫表型 

分 类 号：R733[医药卫生—肿瘤]