人脑胶质瘤细胞裸鼠原位移植双荧光模型的建立  

Establishment of double fluorescent animal models by orthotopic transplantation of human glioma cells into nude mice

作　　者：于宁[1] 李张昱 高鑫[1] 郭剑锋[1] 陈四方[1] 朱宏伟[1] 叶永造[1] 谭国伟[1] 

机构地区：[1]厦门大学附属第一医院神经外科,厦门361003

出　　处：《中华神经医学杂志》2016年第6期554-557,共4页Chinese Journal of Neuromedicine

基　　金：厦门市科技惠民计划项目（2013S0001）

摘　　要：目的建立裸鼠的人脑胶质瘤细胞原位移植双色荧光模型。方法体外培养转染有增强型红色荧光蛋白（ERFP）的人U87细胞（ERFP—U87），制成细胞悬液后接种于表达增强型绿色荧光蛋白（EGFP）的裸小鼠脑部右侧尾状核区域，活体成像系统上动态观察肿瘤的生长，4周后HE染色移植瘤组织切片，荧光显微镜下观察肿瘤细胞与宿主基质细胞的关系。结果表达EGFP的裸鼠在活体成像系统下可见全身组织器官均带有绿色荧光。接种ERFP-U87细胞后1周于移植部位可见红色荧光，且随时间的增加而增强。HE染色移植瘤组织显示肿瘤细胞较正常细胞深染，核大，可见分裂相及多核；荧光显微镜下观察可见肿瘤组织与宿主基质分别呈现红、绿色荧光，肿瘤组织与鼠脑组织界限不清，可见肿瘤发生血管，正常内皮细胞形成血管，部分管壁有肿瘤细胞包绕，肿瘤组织中呈现绿色荧光。结论双色荧光胶质瘤动物模型可作为胶质瘤基础研究的理想模型。Objective To establish the double fluorescent animal models of orthotopic transplantation of human glioma cells into nude mice and provide dynamic observation of occurrence and development ofgliomas. Methods Human U87 cells were transfected with enhanced red fluorescent protein （ERFP） and then inoculated into the right caudate nucleus of nude mice with enhanced green fluorescent protein （EGFP）. Dynamic observation of tumor growth was carried out by in vivo imaging. HE staining was performed on the transplanted tumor tissues 4 weeks after inoculation; the relation of tumor cells and host matrix cells was observed under fluorescence microscope. Results Green fluorescence could be seen in the whole body tissues and organs of nude mice expressed EGFP by in vivo imaging. Red fluorescence could be noted at the graft sites at one week after ERFP-U87 inoculation, and the red fluorescence enhanced as time prolonged. HE staining indicated that the tumor cells in the transplanted tumor tissues were deep dyed with big nucleus; these cells with multinuclear were at division phase. Fluorescence microscope showed that tumor cells and host matrix cells enjoyed red fluorescence and green fluorescence, respectively. Boundary between tumor tissues and mice brain tissues was not clear, tumorigenesis vessels could be seen; normal endothelial cells formed the vessels and some tube walls had tumor cells. Conclusion In these animal models, tumor growth can be dynamically observed, thus, they may serve as ideal models in basic research on gliomas.

关 键 词：神经胶质瘤 原位移植模型 活体成像 绿色荧光蛋白 红色荧光蛋白 

分 类 号：R739.4[医药卫生—肿瘤] R-332[医药卫生—临床医学]