人脑胶质瘤细胞裸鼠原位移植动物模型的建立

Experimental study on the establishment for orthotopic implantation model of human glioma cells in nude mice

目的探讨人脑胶质瘤细胞裸鼠原位移植动物模型建立的技术方法。方法借助动物立体定向仪的引导,采用微注射方法将体外培养人脑胶质瘤细胞U87MG(悬浮于无血清RPMI 1640培养液中,细胞数为108/ml)接种于裸鼠(BALB/c)额叶白质区。接种后观察不同种实验鼠的生存状态,分别于接种后1 h至63 d的不同时间进行裸鼠脑肿瘤组织病理学检查和免疫组织化学分析。结果裸鼠脑内注射体外培养的人脑胶质瘤细胞U87MG的合适速率为0.05μl/min =1μl/20 min,细胞悬液体积1μl,细胞数105,注射时间20 min。按此方案注射U87MG细胞无沿针道返流,恰好在尾状核区形成近圆球形肿瘤体,成瘤率高(100%),肿瘤颅内生长稳定,组织病理学检查符合人脑胶质瘤形态特征,未见脑外转移。结论本研究的人脑胶质瘤细胞裸鼠原位移植动物模型建立方法精确可靠,重复性好。肿瘤符合人脑胶质瘤的生物学特性,该动物模型可作为研究人脑胶质瘤发生、生物学特性以及各种治疗评价的可靠动物模型。

Objective To explore the establishment for orthotopic implantation model of human glioma cells in nude mice. Methods The human brain glioma cell line U87 MG was cultured in vitro, and cells （resuspended in serum-free RPMI 1640 medium at 10^8/ml ） were inoculated by injection with stereotactic guidance into the frontal white matter of nude mice. General observation was conducted after injection. The animals were sacrified at intervals from 1 h to 63 d after implantation and the brain were examined by pathology and immtmohistoehemistry and tumor size was measured. Results Optimal injection rate was fotmd to be 0. 05 μl/min = 1 μl/20 min, a volume of 1μl containing 10^5 cells and injection was done over 20 min. Tumor cells formed a nice sphere in the caudate putamen, away from ventricle as well as brain surface with no noticeable reflux into the needle track. Gliomas were formed in 100% nude mice. And the growth of tumors was stable in the regions of intracerebral environment in which tumors showed the morphological features of human glioma. Conclusion The method allows the highly accurate and reproducible introduction of a given number of cells into a specific area of the nude mice brain. The tumor pathological characteristics mimic human brain glioma. The glioma model in nude mice is reliable animal model for the study of the tumorigenesis, biological characteristics and the evaluation of various treatment medalities.