天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2013年
8期
759-762
,共4页
李新灵%王志爽%邵晓枫%任峰
李新靈%王誌爽%邵曉楓%任峰
리신령%왕지상%소효풍%임봉
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%乳腺肿瘤%肝肿瘤%细胞因子诱导的杀伤细胞
免疫錶型分型%細胞,培養的%細胞繫,腫瘤%白血病%淋巴瘤%肺腫瘤%乳腺腫瘤%肝腫瘤%細胞因子誘導的殺傷細胞
면역표형분형%세포,배양적%세포계,종류%백혈병%림파류%폐종류%유선종류%간종류%세포인자유도적살상세포
immunophenotyping%cells,cultured%cell line,tumor%leukemia%lymphoma%lung neoplasms%breast neo-plasms%liver neoplasms%cytokine-induced killer cells
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。
目的體外誘導培養細胞因子誘導的殺傷性細胞（CIK），研究其增殖、免疫錶型變化及對不同腫瘤細胞繫的殺傷作用。方法從外週血中分離單箇覈細胞，多種細胞因子進行誘導，擴增培養CIK細胞，細胞計數檢測其增殖能力，流式細胞儀檢測細胞免疫錶型，MTT法檢測CIK細胞對人紅白血病細胞繫K562、人B淋巴瘤細胞繫BJAB、人肺癌細胞繫A549、人乳腺癌細胞繫MCF-7、人肝癌細胞繫HepG2的細胞毒性作用。結果 CIK細胞在第5天開始進入快速增殖階段，20 d後增殖為原種植細胞數的182倍，免疫錶型為CD3+（97.83±1.03）%、CD3+CD8+（77.12±1.60）%、CD3+CD56+（27.58±2.02）%。CD3+、CD3+CD8+、CD3+CD56+的比例隨著培養時間的延長增殖迅速（P＜0.01）；培養第13天，細胞殺傷效靶比為40∶1的情況下對細胞繫進行殺傷，K562為（88.89±7.22）%、BJAB為（75.42±9.52）%、A549為（63.19±5.67）%、MCF-7為（43.53±6.31）%、HepG2為（42.63±7.69）%，CIK細胞對以上細胞繫均有明顯的殺傷活性。結論通過細胞因子的誘導培養，可以在體外大量增殖CIK細胞，CIK細胞對不同腫瘤細胞株有很彊的殺傷作用，具有很高的臨床應用價值。
목적체외유도배양세포인자유도적살상성세포（CIK），연구기증식、면역표형변화급대불동종류세포계적살상작용。방법종외주혈중분리단개핵세포，다충세포인자진행유도，확증배양CIK세포，세포계수검측기증식능력，류식세포의검측세포면역표형，MTT법검측CIK세포대인홍백혈병세포계K562、인B림파류세포계BJAB、인폐암세포계A549、인유선암세포계MCF-7、인간암세포계HepG2적세포독성작용。결과 CIK세포재제5천개시진입쾌속증식계단，20 d후증식위원충식세포수적182배，면역표형위CD3+（97.83±1.03）%、CD3+CD8+（77.12±1.60）%、CD3+CD56+（27.58±2.02）%。CD3+、CD3+CD8+、CD3+CD56+적비례수착배양시간적연장증식신속（P＜0.01）；배양제13천，세포살상효파비위40∶1적정황하대세포계진행살상，K562위（88.89±7.22）%、BJAB위（75.42±9.52）%、A549위（63.19±5.67）%、MCF-7위（43.53±6.31）%、HepG2위（42.63±7.69）%，CIK세포대이상세포계균유명현적살상활성。결론통과세포인자적유도배양，가이재체외대량증식CIK세포，CIK세포대불동종류세포주유흔강적살상작용，구유흔고적림상응용개치。
Objective To investigate the proliferation, immune phenotype and cytotoxicity on different cell lines of cytokine-induced killer (CIK) cells collected from healthy donors. Methods Peripheral blood mononuclear cells (PBMC) from healthy donors were induced to become CIK cells by adding cytokines including rhIL-2, rhIFN-γand CD3 McAb. The 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 and HepG2, was detected by MTT assays on day 13. Results CIK cells quickly proliferated from day 5, and expanded by 182-fold after 20-day culture. The immunophenotypes of CD3+, CD3+CD8+and CD3+CD56+were (97.83±1.03)%, (77.12±1.60)%and (27.58± 2.02)%. The percentages of CD3+, CD3+CD8+and CD3+CD56+increased noticeably (P<0.01). According to the effector-tar-get ratio of 40∶1, the activity of CIK cells against tumor cells K562, BJAB, A549, MCF-7 and HepG2 were (88.89±7.22)%, (75.42±9.52)%, (63.19±5.67)%, (43.53±5.67)%and (42.63±7.69)%. The experiments showed that CIK cells possessed high-er antitumor cytotoxic activity. Conclusion CIK cells can be largely capacity cultured by adding cytokines in vitro. CIK cells were a highly efficient cytotoxic cell against tumors, and had clinical application potentials.
