不同激活方式影响细胞因子诱导杀伤细胞的扩增与活性

目的：研究不同激活方式对CIK细胞(Cytoline-induced killer cells，CIK)体外扩增的影响。方法：使用植物血球凝集素(pHYTOHEMAGGLUTININ，PHA)或抗CD3单抗刺激细胞增殖，从脐血单个核细胞定向诱导CIK细胞，用乳酸脱氢酶(LDH)分析法分析细胞毒活性，用流式细胞分析法对不同培养条件下CIK细胞的纯度进行分析。结果:使用抗CD3 mAb刺激生成的CIK细胞在扩增能力、纯度和细胞毒性方面都优于使用PHA刺激生成的CIK细胞。在细胞因子和有丝分裂原加入3d后，将其洗脱，但仍加入IL-2，有利于提高细胞的扩增能力。另外，使用包被的方法将抗CD3 mAb固定在孔板上比悬浮加入效果好。结论：这一结果对于CIK细胞的大量扩增和临床应用具有指导意义。

Effect of various activation ways on the in vitro proliferation of CIK cell

Objective To study various activation ways on the in vitro proliferation of CIK cell. Methods PHA and CD3 monoclonal antibody were employed to enhance the cell in vitro replication. CIK cells were induced from mononuclear cells (MNC) which were separated from human cord blood and evaluating lactate dehydrogenase release for the cytotoxicity assay and flow cytometer for cell purity analy- sis were used in this study. Results It was found that CD3 monoclonal antibody was more favorable for the CIK than PHA in terms of cell proliferation, purity and cytotoxicity. It was also observed that more fold of cell replication could be obtained when cytokines and mitogenic stimu- lator were washed out after three days but still adding IL-2. What's more, better results were achieved when CD3 monoclonal antibody was immobilized in the multi-well plate compared with the case that it was in the medium. Conclusion This work is important for the CIK in vitro pro- liferation and clinical application in the future.