人脑胶质瘤干细胞移植于表达绿色荧光蛋白裸小鼠的研究  

Transplantation of human glionm stem cells in nude mice with green fluorescent protein expression

作　　者：吴自成[1] 黄强[1] 邵义祥[2] 薛智谋[3] 董军[1] 刁艺[1] 王爱东[1] 兰青[1] 

机构地区：[1]苏州大学附属第二医院神经外科,215004 [2]南通大学实验动物中心 [3]苏州大学实验动物中心

出　　处：《中华医学杂志》2008年第33期2317-2320,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目（30672164,30772241）;江苏省自然科学基金资助项目（BK2007507）

摘　　要：目的培育表达绿色荧光蛋白（GFP）的裸小鼠，并探讨将其用于人胶质瘤干细胞（HGSC）移植实验研究。方法将C57BL／6J-GFP转基因小鼠与NC系裸小鼠进行交配，在继代时严格挑选都表达GFP的无毛雄鼠与有毛母鼠交配，通过肉眼、免疫组织化学和荧光显微镜等方法观察GFP在裸小鼠皮肤和脏器中的表达情况，继将HGSC原位移植于表达GFP的裸小鼠，以观察移植瘤的生长情况。结果传至8代的裸小鼠，包括脑在内的全身主要器官和细胞都表达GFP。对用于HGSC原位移植的脑，能清晰辨认出不发光的瘤细胞和发光的宿主细胞。而瘤细胞经人特异白细胞抗体（HLA）连接红色荧光剂（PE）的免疫组织化学染色后，在共聚焦显微镜下可清晰显示与宿主脑细胞的组织学关系。结论用免疫功能健全、表达GFP的转基因鼠与免疫功能缺陷的裸小鼠杂交，可获得脑等组织器官表达GFP的裸小鼠。对其进行HGSC原位移植，因可清晰辨认移植瘤与宿主组织的关系而有望用于肿瘤组织重构的研究。Objectives To investigate the possibility of transplantation of human glioma stem cells （HGSCs） in nude mice stably expressing green fluorescent protein （GFP） so as to clearly identify the incubated HGSCs from the host tissues. Methods Transgenic C57BL/6J mice expressing GFP was crossed with nude mice of the line NC, then hairless male nude mice expressing GFP were crossed with hairy female pubescent mice to obtain nude mice with GFP expression the expression of GFP in the skin and organs of these nude mice were evaluated by naked eyes, and immunohistochemical and immunofluorescence assays. HGSCs were transplanted orthotopically into the caudate nuclei of nude mice expressing GFP. Immunohistochemistry was used to observe the transplanted tumor. Results The structures rich in adipose tissue of the 8th generation nude mice were dark green and the other organs were light green. However, green fluorescence was emitted from all tissues under fluorescence microscopy. Confocal fluorescence microscopy showed that the tumor cells were stained red, distinguished from the host cells distinctly in the brains bearing tumor transplanted orthotopically. Conclusion Nude mice expressing GFP can be obtained by crossing the trangenic mice bearing naive immunity with nude mice. Orthotopic transplantation of HGSCs may be used in the investigation of tumor tissue reconstitution because of the easy identification between the transplantation tumor and host tissue.

关 键 词：神经胶质瘤 绿色荧光蛋白 转基因小鼠 肿瘤移植 肿瘤干细胞 

分 类 号：R686[医药卫生—骨科学]