绿色荧光裸鼠的建立和验证  

Establish and Verification of Transgenic EGFP Nude Mouse

作　　者：闫寒[1] 贺晓玉[1] 秦超[1] 李小颖[1] 高虹[1] 

机构地区：[1]中国医学科学院,北京协和医学院,医学实验动物研究所,卫生部人类疾病比较医学重点实验室,国家中医药管理局人类疾病动物模型三级实验室,北京100021

出　　处：《中国比较医学杂志》2012年第6期62-64,86,F0003,共5页Chinese Journal of Comparative Medicine

基　　金：重大新药创制科技重大专项课题(2008ZX09305-001)

摘　　要：目的建立系统性表达绿色荧光蛋白的裸鼠,接种人源肺癌细胞验证该模型是否具有免疫缺陷性,并观察双色荧光的成像效果。方法利用系统性表达绿色荧光蛋白的C57BL/6J小鼠与BALB/C裸小鼠多代杂交和互交,建立稳定表达绿色荧光蛋白的裸鼠。大体解剖观察胸腺生长情况,整体和器官荧光成像验证绿色荧光蛋白的表达情况。以2×106/只的剂量对其皮下腋下接种表达红色荧光蛋白的人类A549肺癌细胞(RFP-A549),通过观测肿瘤生长来验证模型的免疫缺陷性。同时,利用红色荧光标记的肿瘤和绿色宿主鼠,对双色的整体成像效果进行观测。结果构建出系统性表达绿色荧光蛋白的裸鼠,大体解剖可见胸腺缺失。在激发光的激发下,绿色荧光裸鼠全身发出清晰的绿色荧光,脑、心脏、肺脏、肝脏、肾脏,肠胃及胰腺等主要器官可见明显绿色荧光。接种RFP-A549细胞后,成瘤率达到100%,整体动物荧光成像表现出清晰的双色。结论本研究构建出的绿色荧光裸鼠。Objective To develop a transgenic GFP nude mouse model,human lung carcinoma cells were implanted to test and verify the Immunodeficiency of the model,then observing the result of Dual-color fluorescence imaging.Method The GFP nude mouse was obtained by crossing BALB/c male nude mice with the transgenic EGFP female C57/B6 mouse.We observe the Thymus through Gross Anatomy,test GFP expression Through Whole-Body and Major Organs Fluorescence Imaging.Six-week-old nude EGFP mouse was injected RFP-expressing Human A549 lung carcinoma cells（RFP-A549） subcutaneously with a single dose of 2×106,test and verify the Immunodeficiency of EGFP-expressing nude mice through The development of tumor.Meanwhile,a GFP mouse-RFP tumor model was established to evaluate the Dual-color fluorescence imaging.Result Developed EGFP-expressing nude mice,obvious fluorescence can be observed in whole-body and major organs include brain,heart,lungs,liver,kidney,intestines,stomach and pancreas.After implanted RFP-A549 cells subcutaneously,the percentage of tumorigenesis is 90%,fluorescence imaging show dual-color clearly.Conclusion Developed a transgenic GFP nude mouse model,the whole-body fluorescence imaging clearly and have Immunodeficiency.

关 键 词：绿色荧光裸鼠 RFP-A549细胞 双色荧光成像 

分 类 号：R332[医药卫生—人体生理学]