大体系冻存对细胞因子诱导的杀伤细胞免疫表型及细胞杀伤活性的影响

摘要

Objective Effect of large system (50 mL) cryopreservation on immunophenotype and killing activities of cytokine induced killer (CIK) cells.Methods Peripheral blood samples were collected from 6 healthy volunteers and then the mononuclearcell were isolated by Ficoll method and were induced to CIK cells.The CIK cells were reanimated at 1 month after cryopreservation and collected.The cell survival rate and immunophenotype was detected by flow cytometry before and after cryopreservation.The killing activity of CIK cells was detected by culturing with breast cancer cells (target ratio 10 ∶ 1 and 40 ∶ 1) and compared with fresh cell;and the killing activity of CIK cells under different target ratio was compared between before and after cryopreservation.Results The average cell survival rate of CIK cells before cryopreservation was (97.79 ± 1.92) ％,which was significantly higher than that after reanimation(83.61 ± 3.42) ％ (P ＜ 0.05).The proportion of CIK with CD3 +,CD3 + CD4 +,CD3 + CD8 +,CD3 + CD56 + phenotype after reanimation was slightly lower than that before cryopreservation,but the difference was not statistically significant(P ＞ 0.05).There was no significant difference in the cell killing rate of CIK cells before cryopreservation and after reanimation when the target ratio was 10 ∶ 1 and 40 ∶ 1 (P ＞0.05);the cell killing rates of CIK cells before cryopreservation and after reanimation when the target ratio was 40 ∶ 1 were significantly higher than those when the target ratio was 10 ∶ 1 (P ＜ 0.05).Conclusion The large system (50 mL) cryopreservation can effect the survival rate of CIK cells,but has no effect on the immune phenotype and cell activity.%目的 探讨大体系(50 mL)冻存对细胞因子诱导的杀伤(CIK)细胞免疫表型及细胞杀伤活性的影响.方法 采集6例健康志愿者的外周血,采用Ficoll法分离得到单个核细胞,用细胞因子诱导培养CIK细胞.收集冻存1个月后复苏的CIK细胞,采用流式细胞术检测其冻存前后的细胞活率和免疫表型;通过与乳腺癌细胞共培养(效靶比10∶1和40∶1)的方法测定其杀伤活性,与新鲜未冻存的细胞杀伤活性进行比较,同时并对不同效靶比冻存前后的细胞杀伤活性进行比较.结果 CIK细胞大体系冻存前平均细胞活率(97.79 ± 1.92)％,显著高于复苏后的(83.61±3.42)％(P＜0.05).复苏后CIK细胞中CD3+、CD3+ CD4+、CD3+ CD8+、CD3+ CD56+表型CIK细胞所占比例虽较冻存前稍有降低,但差异均无统计学意义(P＞0.05).效靶比10∶1、40∶1时冻存前细胞和复苏后细胞杀伤率比较差异均无统计学意义(P＞0.05);效靶比40∶1时冻存前细胞和复苏后细胞杀伤率均显著高于效靶比10∶1时(P＜0.05).结论 大体系(50 mL)冻存对于CIK细胞活率虽有影响,但对免疫表型和细胞活性均未影响.