摘要
目的采用活细胞荧光成像技术动态观察胶质瘤干细胞向内皮细胞的分化过程,并分析细胞表面标记物的变化。方法以胶质瘤干细胞株GSC-1为对象,分别进行免疫荧光标记技术和活细胞荧光成像技术观察其在分化过程中分子标记物的变化趋势。结果 GSC-1置入三维培养基质,细胞随时间增加而增殖,6 h后,细胞突起发生融合,细胞增殖,可清晰地观察到多细胞所形成的圆形、多边形和三角形管道样结构。VEGF诱导下免疫荧光染色观察GSC-1表达CD34随时间增加而增加,6 h时较多GSC-l细胞表达CD34,出现典型的闭合样管腔结构。CD34阳性细胞数间差异存在统计学意义(P<0.05)。活细胞荧光观察追踪单个细胞发现,6 h达到高峰,CD34阴性细胞逐渐出现绿色荧光,提示转变成CD34阳性细胞。结论活细胞荧光成像技术观察结果同传统免疫荧光技术所得到的结果相一致,同时可获得单个细胞从CD34阴性细胞转变为CD34阳性细胞的直接影像证据,可直观、准确地对单个细胞进行追踪。
Objective To observe the differentiation of glioma stem cells into endothelial cells by living cell fluorescence imaging, and to analyze the changes of cell surface markers. Methods Glioma stem cells line GSC - 1 were used as the target. Immunofluorescence labeling technique and living cell fluorescence imaging tech-nique were used to observe the changes of molecular markers during the differentiation. Results GSC - 1 cells were implanted into three dimensional culture medium, and the cells proliferated with time. After 6 h, the cell processes fused and proliferated, and the circular - , polygonal - and triangular tube - like structures were clearly observed. Immunofluorescence staining showed that the expression of CD34 in GSC - 1 increased with time, and the CD34 was expressed by many cells in 6 h. There were typical closed lumen - like structures in VEGF cells. The difference of CD34 positive cells was statistically significant （P 〈0. 05）. Single cell was observed by living cell fluorescence observation. After 6 h,the intensity of the fluorescence reached the peak, and CD34 negative cells gradually showed green fluorescence, suggesting that the cells were transformed into CD34 positive cells. Conclusion The results of living cell fluorescence imaging are consistent with the results obtained by traditional immunofluorescence technique. Simultaneously, the direct imaging evidence of single cell transforms from CD34 negative cells into CD34 positive cells can be obtained, which can directly and accurately track individual cells.
作者
魏强国
王浩
刘欣民
WEI Qiangguo;WANG Hao;LIU Xinmin(Department of Neurosurgery, Nanshan District peoples Hospital of Shenzhen （Shenzhen, 518052)
出处
《生物医学工程学进展》
CAS
2017年第4期196-198,共3页
Progress in Biomedical Engineering
关键词
活细胞荧光成像技术
胶质瘤干细胞
内皮细胞
分化
分子标记物
living cell fluorescence imaging technique,glioma stem cells,endothelial cells,differentia- tion, molecular markers
