绿色荧光裸鼠的建立和验证

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C57 BL/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,动物整体可以清晰地表达绿色荧光并具有免疫缺陷性%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 ×10 , 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.