CIK在无血清培养体系中的增殖、表型变化和抗肿瘤活性

The proliferation, phenotype change and anti-tumor activity of cytokine induced killer cells

目的:探索细胞因子诱导的杀伤细胞(CIK)在无血清体系中的增殖规律、表型变化及其抗瘤活性。方法:不同培养基培养CIK细胞,采用活细胞计数法观察CIK细胞的增殖,流式细胞仪检测CIK细胞的表型,CytoTox96非放射性细胞毒试剂盒检测CIK细胞的细胞毒活性。结果:经过细胞因子和抗体刺激后,CIK细胞能明显增殖,无血清培养基加自体血浆组最高可扩增473.28±27.53倍,无血清培养基组可扩增218.24±16.86倍,而RPMI1640加FCS只扩增11.52±1.04倍。CD3++CD8+、CD3++CD56+、CD226++CD11a+和CD305++CD11a+细胞随着培养时间的延长而增加,而CD3++CD4+细胞则明显减少。CIK细胞对肿瘤细胞的细胞毒作用明显高于LAK细胞(P<0.01),且其细胞毒活性随着培养时间的延长而增高。结论:CIK细胞在体外扩增能力强,对肿瘤细胞的杀伤活性高,有望成为新一代抗肿瘤过继免疫细胞制剂而应用于临床。

AIM： to study the growth regularity, the phenotype change and the cytotoxicity of CIK cells. METHODS： The number of CIK cells was counted by living cell counting in different culturing time to observe the growth of the CIK cells, and the phenotype change of the CIK cells was detected by flow cytometry. Meanwhile cytotoxicity of CIK cells to tumor cell lines was also detected by CytoTox96 non-radiated cytotoxicity kit. RESULTS： After stimulated by cytokines and anti-CD3 antibody, CIK cells can proliferate significantly. The cell number of CIK was increased to 473.28 ± 27.53 fold in serum-free medium plus auto-serum, 2.18.2.4 ±16. 86 fold in serum-free medium and only 11.52 ± 1.04 fold in RPMI1640 plus fetal FCS, respectively. The CD3^＋ ＋CD8 ^＋ , CD3 ^＋ ＋CD56^＋ , CD226^＋ ＋CD11a^＋ and CD305 ^＋ ＋CD11a^＋ cells were increased with the progression of the cultural time and the CD3^＋ ＋ CD4^＋cells were decreased with the progression of cultural time. The cytotoxicity of CIK cells to tumor cell lines was significantly higher than that of LAK cells （P 〈0.01 ） and its cytotoxicity was increased with progression of the cultural time. CONCLUSION： CIK cells have strong proliferative ability and higher cytotoxicity to tumor cells in vitro, which could be used as a potential antitumor adoptive immunotherapy in clinic.