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目的:利用原子力显微镜(AFM)观测细胞因子诱导的杀伤细胞(CIK)分化过程。方法:流式细胞术检测CIK表型变化,MTT法检测CIK细胞毒活性,免疫磁珠法分离CIK,AFM动态观测CIK细胞表面形态变化。结果:CIK在培养d15～d18数量达到并维持于最高水平。培养d16CIK细胞中CD3+CD8+、CD3+CD56+细胞群较d10均有不同程度的增加,分别占到(65.17±0.11)%和(17.25±0.04)%。成熟的CIK对多种肿瘤细胞均显示了较强的体外杀伤能力,利用AFM观测其细胞膜表面重要指标即平均粗糙度、峰数、平均峰值高度、平均低谷高度、表面积差值均高于培养d10细胞,P<0.01。结论:成熟CIK细胞膜表面生物蛋白分子明显增加,其细胞膜形态和功能变化密切相关。
OBJECTIVE: To observe the differentiation of cytokine-induced killer (CIK) cells by atomic force microscopy (AFM). Methods: The phenotypic changes of CIK cells were detected by flow cytometry. The cytotoxicity of CIK cells was detected by MTT assay. CIK was separated by immunomagnetic beads method. The morphological changes of CIK cells were observed by AFM. Results: The number of CIK reached and maintained the highest level in d15 ~ d18. The cell population of CD3 + CD8 + and CD3 + CD56 + in d16CIK cells increased more than d10, which accounted for (65.17 ± 0.11)% and (17.25 ± 0.04)%, respectively. Mature CIK showed strong in vitro cytotoxicity against various tumor cells. The average surface roughness, peak number, average peak height, average height of trough and surface area were significantly higher than those of cultured d10 Cells, P <0.01. CONCLUSION: The number of biological protein molecules on the surface of mature CIK cells is significantly increased, and the changes of cell membrane morphology and function are closely related.