细胞因子诱导的自然杀伤细胞培养体系的评价  

Culture system of cytokine-induced natural killer cell

作　　者：王娟[1,2] 桑圣刚[3] 郝新宝[1] 

机构地区：[1]海南医学院附属医院血液肿瘤内科,海南海口571100 [2]贵阳中医学院,贵州贵阳550000 [3]海南医学院检验系,海南海口571100

出　　处：《海南医学院学报》2011年第3期302-304,共3页Journal of Hainan Medical University

基　　金：海南省卫生厅基金项目(2008-61)~~

摘　　要：目的:评价采用γ-干扰素(IFNγ-)、白介素-2(IL-2)、抗CD3单抗3因子对外周血来源的CIK细胞培养的效果。方法:采集17例实体肿瘤患者的外周血,分离单个核细胞培养,于培养第1天加入重组人IFN-γ,第2天加入抗人CD3单抗、重组人IL-2,隔天半量换培养液并添加人重组IL-2以维持其浓度,培养21 d收获细胞。于培养第1、7、14、21天进行细胞计数,同时于培养第1、8、14、21天测定细胞免疫表型。结果:细胞因子共刺激2～3 d即出现细胞集落,14 d后CIK细胞总数达(6～17)×109/L,较培养前扩增47～150倍,细胞存活率大于95%;具有免疫表型CD3+、CD4+、CD8+和CD3+CD56+细胞的平均值分别从培养前的(65.1±1.9)%、(36.9±0.8)%、(28.5±3.1)%、(1.8±0.4)%增加到培养第21天的(89.6±2.1)%、(50.1±1.9)%、(57.2±2.5)%、(36.1±4.2)%。结论:按照上述方法培养的CIK细胞在体外扩增率高,方法可行,CIK细胞在体外培养到14～21 d,CD3+CD56+呈现高表达,此时期适宜应用于临床治疗,且患者自体外周血易获得、整个培养过程简便,值得临床应用推广。Objective：To evaluate culture effects of cytokine-induced natural killer cell by IFN-γ,IL-2,anti-CD3,monoclonal antibody factor 3.Methods：Mononuclear cells were separated from peripheral blood that were collected from 17 patients with tumor and cultured.IFN-γ was added on day 1;IL-2 and anti-CD3 were added on day 2.Then 0.5 day later,half the amount of medium was abandoned,but the concentration was maintained by adding IL-2.The cultured cells were harvested on day 21.Cell amount were counted on day 1,7,14 and 21,and immunophenotyping detected on day 1,8,14,and 21.Results：After 2-3 days of culture,cell colony appeared.After 14 days,total number of CIK cells was（6～17）×109/L,47-150 times more than the original samples.More than 95% cells survived;average number of cells that showed immunophenotypes of CD3＋,CD4＋,CD8＋,CD3＋ CD56＋ before and 21 days after the culture were（65.1±1.9）%,（36.9±0.8）%,（28.5±3.1）%,（1.8±0.4）% vs（89.6±2.1）%,（50.1±1.9）%,（57.2±2.5）%,（36.1.1±4.2）%.Conclusions：By the above mentioned culture system,number of cytokine-induced natural killer can be increased quickly,and it shows high expression of CD3＋,and CD56＋ showed high expression.Since autologous peripheral blood of patients is easy to be collected and this method is worthy of clinical application.

关 键 词：细胞因子诱导的自然杀伤细胞 细胞培养 Γ-干扰素 白介素-2 抗CD3单抗 

分 类 号：Q26[生物学—细胞生物学] R392.1[医药卫生—免疫学]