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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。
Objective: To explore the proliferation, phenotypic changes and antitumor activity of cytokine-induced killer cells (CIK) in serum-free system. Methods: CIK cells were cultured in different media. The proliferation of CIK cells was observed by viable cell counting. The phenotype of CIK cells was detected by flow cytometry. The cytotoxic activity of CIK cells was detected by CytoTox96 non-radioactive cytotoxic kit. Results: CIK cells proliferated significantly after stimulated by cytokines and antibodies. The highest proliferation of CIK cells was 473.28 ± 27.53 times in serum-free medium plus 218.24 ± 16.86 times in serum-free medium, while RPMI1640 plus FCS Only amplified 11.52 ± 1.04 times. CD3 ++ CD8 +, CD3 ++ CD56 +, CD226 ++ CD11a + and CD305 ++ CD11a + cells increased with incubation time, whereas CD3 ++ CD4 + cells decreased significantly. The cytotoxic effect of CIK cells on tumor cells was significantly higher than that of LAK cells (P <0.01), and the cytotoxic activity of CIK cells increased with the prolongation of culture time. CONCLUSION: CIK cells have strong ability to expand in vitro and high cytotoxic activity against tumor cells, which is expected to be a new generation of anti-tumor adoptive immune cell preparations for clinical application.