细胞因子诱导杀伤细胞分化过程中的原子力显微镜观察

Observation of cytokine-induced killer cells during the differentiation process using atomic force microscopy

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 process of differentiation of CIK （cytokine-induced killer） using the atomic force microscopy.METHODS：The phenotype of CIK was detected by flow cytometry,the cytotoxicity was tested with MTT method and the CIK cells were separated by immunomagnetic beads.Moreover,the morphologic changes on CIK membranes were observed by AFM （atomic force microscopy）.RESULTS：Between the 15 th and 18th day of culture,the number of CIK reached and maintained the highest level.On the 16th day,the CD3＋CD8＋ and CD3＋CD56＋ cells of CIK increased compared with the 10th day,which accounted to （65.17±0.11）% and （17.25±0.04）%.Mature CIK showed powerful abilities to kill and wound various kinds of tumor cells in vitro and the important AFM parameters of mean roughness,peak count,average max peak height,average max depth and surface area difference were significantly higher than those of the cells on the 10th day （P0.01）.CONCLUSION：The number of membrane bioprotein on mature CIK increases intensively,and the morphous of its membrane is intimately correlated with the functional changes.