天津医药

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CIK细胞体外诱导培养及对不同肿瘤细胞株的细胞毒作用

李新灵,王志爽,邵晓枫,任峰   

  1. 天津斯坦姆再生医学技术研究开发中心
  • 收稿日期:2012-11-02 修回日期:2013-01-09 出版日期:2013-08-15 发布日期:2013-08-15
  • 通讯作者: 李新灵

Effects of Cytokine-Induced Killer Cells on Different Tumor Cell Lines in Vitro

LI Xinling ,WANG Zhishuang ,SHAO Xiaofeng ,REN Feng   

  1. Tianjin STEM Regenerative Medicine Technical Research Development Center, Tianjin 300384, China
  • Received:2012-11-02 Revised:2013-01-09 Published:2013-08-15 Online:2013-08-15
  • Contact: LI Xinling

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摘要：

【摘要】 目的 体外诱导培养细胞因子诱导的杀伤性细胞（CIK）， 研究其增殖、免疫表型变化及对不同肿瘤细胞系的杀伤作用。 方法 从外周血中分离单个核细胞， 多种细胞因子进行诱导， 扩增培养CIK细胞， 细胞计数检测其增殖能力， 流式细胞仪检测细胞免疫表型， MTT法检测 CIK细胞对人红白血病细胞系 K562、人B 淋巴瘤细胞系BJAB、人肺癌细胞系 A549、人乳腺癌细胞系 MCF-7、人肝癌细胞系 HepG2的细胞毒性作用。 结果 CIK细胞在第 5 天开始进入快速增殖阶段， 20 d 后增殖为原种植细胞数的 182 倍， 免疫表型为 CD3 +（97.83±1.03）%、CD3 +CD8 +（77.12±1.60）%、CD3+CD56（ + 27.58±2.02）%。 CD3+、CD3+CD8+、CD3+CD56+的比例随着培养时间的延长增殖迅速（P＜0.01）； 培养第 13 天， 细胞杀伤效靶比为 40∶ 1 的情况下对细胞系进行杀伤， K562 为（88.89±7.22）%、BJAB 为（75.42±9.52）%、A549 为（63.19±5.67）%、MCF-7 为（43.53±6.31）%、HepG2 为（42.63±7.69）%， CIK细胞对以上细胞系均有明显的杀伤活性。 结论 通过细胞因子的诱导培养， 可以在体外大量增殖 CIK细胞,CIK细胞对不同肿瘤细胞株有很强的杀伤作用， 具有很高的临床应用价值。

关键词: 免疫表型分型, 细胞, 培养的, 细胞系, 肿瘤, 白血病, 淋巴瘤, 肺肿瘤

Abstract: Objective   To investigate the proliferation, immune phenotype and cytotoxicity on different cell lines of CIK cells collected from healthy donors. Methods   Peripheral blood mononuclear cell(PBMC) from healthy donors were induced to become CIK cells by adding cytokines including rhIL-2, rhIFN-γ and CD3 McAb. Proliferation of CIK cells was tested by blood cell recording board. The CIK cells were analyzed on different time points by FACS; The cytotoxicity of CIK cells against different tumor cell lines such as K562, BJAB, A549, MCF-7, HepG2 in vitro by MTT assays on day 13. Results  CIK cells quickly proliferated from day 5, and expanded by 182-fold after 20 days’ culture. The immunophenotypes of CD3+, CD3+CD8+, CD3+CD56+ were (97.83±1.03) %, (77.12±1.60) % and (27.58±2.02) %. The percentages of CD3+, CD3+CD8+, CD3+CD56+ increased noticeably (P<0.01). According to the effector-target ratio of 40:1, the activity of CIK cells against tumor cells K562, BJAB, A549, MCF-7, HepG2 were (88.89±7.22) %, (75.42±9.52) %, (63.19±5.67) %, (43.53±5.67) % and (42.63±7.69) %. The experiment showed that CIK cells possessed higher antitumor cytotoxic activity. Conclusion   CIK cells can be largely capacity cultured by adding cytokines in vitro. CIK cells was a highly efficient cytotoxic cells against tumors, and had clinical application potentials.

Key words: Immunophenotyping, cells, cultured, cell line, tumor, leukemia, lymphoma, 肺肿瘤