CFSE荧光负染法检测RAW264.7小鼠单核巨噬细胞白血病细胞摄取超顺磁性氧化铁纳米粒子  

In vivo Cell CFSE Fluorescence Negative Staining for Detection of Super Paramagnetic Iron Oxide Nanoparticles Phagocytosed by Mouse Mononuclear Macrophage Leukemia Cells——RAW264.7

作　　者：何向锋[1,2] 杨俐萍[3] 陈宝安[1] 王建红[2] 文颂[4] 施文[5] 徐薇薇[2] 凌国杰 

机构地区：[1]东南大学医学院附属中大医院血液科(江苏省医学重点学科),江苏南京230009 [2]南通大学附属肿瘤医院内科,江苏南通226361 [3]南通大学附属肿瘤医院中心实验室,江苏南通226361 [4]南昌大学第二附属医院介入室,江西南昌330006 [5]南通大学附属第二医院普通外科,江苏南通322600

出　　处：《中国实验血液学杂志》2015年第4期1168-1172,共5页Journal of Experimental Hematology

基　　金：国家重点基础研究发展计划(973计划)子课题(2010CB732404);国家高技术研究发展计划(863计划)子课题(2012AA022703);国家自然科学基金(81170492;81370673;81401460);江苏省科技厅医学重点项目(BL2014078);中国博士后科学基金项目(2013M530227);江苏省博士后科研资助计划项目(1301099C);南通市市级科技计划项目(B K2012067)

摘　　要：目的:探讨CFSE荧光负染法用于检测RAW264.7小鼠单核巨噬细胞白血病细胞吞噬超顺磁性氧化铁纳米粒子(super paramaganetic iron oxide nanoparticles,SPIO)的可行性及其荧光特征。方法:体外对RAW264.7细胞进行SPIO的标记,然后分别采用普鲁士蓝染色方法和CFSE荧光负染法检测RAW264.7细胞对SPIO的摄取情况;用台盼蓝染色法检测CFSE荧光负染细胞的活力;应用激光扫描共聚焦显微镜进行荧光负染法检测RAW264.7细胞对SPIO的摄取。结果:普鲁士蓝将RAW264.7细胞内吞噬的SPIO标记为醒目的蓝色,核固红将细胞核标记为红色,在荧光图像上表现为荧光负染区,呈筛孔状。CFSE荧光负染法与普鲁士蓝染色法的检测结果具有很好的对应性。RAW264.7细胞经过SPIO和CFSE双重标记后,对细胞活力的影响较小,死细胞比例较低。结论:CFSE荧光负染法可用于RAW264.7活细胞吞噬SPIO的检测,同普鲁士蓝染色法具有很好的对应性,两者的检测结果可相互印证。Objective : To explore the feasibility and fluorescence characteristics of CFSE negative staining for in vivo cell imaging of super paramagnetic iron oxide nanoparticles( SPIO) phagocytosed by mouse mononuclear macrophage leukemia cells—RAW264. 7. Me thods: After labeled with SPIO,the RAW264. 7 macrophages were stained with Prussian blue stain and CFSE fluorescence negative stain step by step. Furthermore,trypan blue staining was used to evaluate cell viability of cells which stained with CFSE. At last,laser scanning confocal microscope was used to measure SPIO in cells through CFSE fluorescence negative stain method. Re sults: SPIO within RAW264. 7 macrophages showed blue in Prussian's blue staining,while showed negative area in CFSE negative staining. Good consistencies between Prussian's blue staining and CFSE negative staining were observed. In addition,RAW264. 7 macrophages showed high viability after SPIO / CFSE dual-labeled method,proved by typan stain. Conclusion: The CFSE fluorescence negative staining may be used for detecting SPIO that phagocytosed by RAW264. 7 macrophages and it is showed good consistency that confirmed one another when compared to classic Prussian' blue staining.

关 键 词：CFSE荧光负染法 RAW264.7小鼠 单核巨噬细胞 白血病细胞 超顺磁性氧化铁纳米粒子 

分 类 号：R733.7[医药卫生—肿瘤]