【摘要】 目的:探讨患者冻存外周血干细胞体外诱导CIK细胞的增殖能力、表型特征及其对B细胞淋巴瘤细胞系的杀伤效应。方法:复苏患者冻存外周血干细胞悬液,Ficoll密度梯度方法分离单个核细胞及从健康人新鲜外周血分离单个核细胞,经IFNγ、IL-2、CD3McAb诱导培养获得CIK细胞。细胞计数板观察CIK细胞增殖能力;流式细胞术检测CIK的表型;以CFSE标记靶细胞区分效应细胞,PI染色,FACS检测靶细胞死亡率。结果:患者冻存外周血干细胞来源的CIK细胞较健康人增殖速度慢,最大增殖倍数低,但CD3+CD56+细胞比例与健康人差异无统计学意义,分别为(16.14±8.49)%和(17.52±3.30)%,P=0.744。患者及健康人CIK细胞与相应的诱导培养前单个核细胞相比,杀伤活性明显增强,在效靶比10:1时患者及健康人CIK细胞对Daudi细胞的杀伤活性为(29.23±8.85)%、(20.13±2.36)%,与培养前相比P值分别为0.004、0.001;患者CIK细胞杀伤活性强于健康人,P=0.05。结论:患者冻存外周血干细胞可以诱导生成具有抗肿瘤活性的CIK细胞,为其临床应用提供了实验依据。更多还原

【Abstract】 OBJECTIVE:To explore the proliferation capacity and phenotypic characteristics of CIK cells induced from cryo-preserved peripheral blood progenitor cells(PBPCs) of patients and the fresh PBMCs of healthy donors,and compare their cytotoxicities against B lymphoma cell line in vitro.METHODS:Mononuclear cells were isolated from cryo-preserved PBPCs of the patients and fresh peripheral blood of health donors by Ficoll-hypaque density centrifugation,and then induced by IFNγ,IL-2 and CD3mAb.The proliferation capacity of CIK cells was detected by cell recording board;the phenotypes of CIK cells were analyzed by FACS;target cells were differentiated from effect cells by CFSE dying.The mortality of target cells was determined by FACS.RESULTS:The CIK cells induced from cryo-preserved PBPCs of the patients proliferated much more slowly than those from the the fresh PBMCs of healthy donors,and its maximum proliferation multiple was also lower.But the proportion of CD3+ CD56+ cell showed no significant differences between them,being (16.14±8.49)% and (17.52±3.30)% respectively (P=0.744).The cytotoxicity of patients and health donors’s CIK cells was significantly stronger than that of the uncultured PBMCs (P were 0.004 and 0.001).With the effector-target ratio 10:1,the activities against daudi cell line of the CIK cells of the two groups were (29.23±8.85)% and (20.13±2.36)%,and the activities of patients’s CIK cells were more stronger than those of the health donors (P=0.05).CONCLUSIONS:CIK cells owning strong anti-tumor ability can be induced from the cryo-preserved PBPCs of patients.Our findings provide an experimental basis for new applications of cryo-preserved PBPCs in clinic.更多还原