C6_Wistar大鼠脑胶质瘤活体生物发光成像肿瘤动物模型的构建.docVIP

  C6/Wistar 大鼠脑胶质瘤活体生物发光成像 肿瘤动物模型的构建# 全毅1，黄伟1，邵帅2，金明姬1，邵荣光3，高钟镐1** 5 10 15 20 25 30 35 40 45 （1. 中国医学科学院北京协和医学院药物研究所，北京，100050； 2. 延边大学药学院药剂学教研室，吉林延吉，133000； 3. 中国医学科学院北京协和医学院医药生物技术研究所，北京，100050） 摘要：本文旨在构建 C6/Wistar 大鼠脑胶质瘤活体生物发光成像肿瘤动物模型。首先制备稳 定表达萤火虫荧光素酶的大鼠脑胶质瘤细胞株 C6-Luc，然后采用立体定向手术在 Wistar 大 鼠右侧尾状核接种 DAPI 荧光标记的 C6-Luc 细胞，于接种后 0d、7d、14d、21d 通过 Xenogen 活体动物体内成像系统观察肿瘤生长情况，次日起记录体重、神经学评分，处死后灌流取脑 进行常规 HE 染色、GAFP 分析和 S-100 免疫组化染色。同时进行裸鼠 C6-Luc 成瘤实验， 判定细胞体内生长的稳定性。 结果表明肿瘤模型于接种后 7d 出现明显体重及行为改变，病 理切片可见肿瘤细胞沿神经纤维向周边侵袭，占位明显，肿瘤组织内见多处出血灶。免疫组 化染色显示胶质纤维酸性蛋白 GFAP 及中神经纤维 S-100 蛋白强阳性表达，肿瘤细胞聚集， 血脑屏障严重破坏，并沿侵袭组织向对侧生长。大鼠大部分死于 30d 内。本文构建的大鼠脑 胶质瘤活体生物发光成像肿瘤动物模型比较稳定，病理性质与人脑间质、胶质母细胞瘤具有 相似性，可用于抗肿瘤药物或基因治疗的临床前期研究。 关键词：脑胶质瘤；动物模型；活体生物发光成像；荧光素酶；Wistar 大鼠 中图分类号：R965.1 Construction of C6/Wistar brain glioma animal model based on in vivo bioluminescence imaging QUAN Yi1, HUANG Wei1, SHAO Shuai2, JI Mingji1, SHAO Rongguang3, GAO Zhonggao1 (1. Institute of Materia Medica, Chinese Academy of Medical Sciences Peking Union Medical college, Beijing, 100050, China; 2. Department of Pharmaceutics, Yanbian University, Yanji 133000, China; 3. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences Peking Union Medical college, Beijing, 100050, China) Abstract: This study aimed at constructing the C6/Wistar brain glioma animal model based on in vivo bioluminescence imaging. A stable rat glioma cell line C6-Luc expressing firefly luciferase was firstly constructed. The DAPI fluorescence labeled C6-Luc cells were then inoculated in the right caudate nucleus of Wistar rats through stereotactic surgery. After inoculation for 0d, 7d, 14d and 21d, the tumor growth was detected by a Xenogen in vivo animal imaging system. The next day record of weight and neurological score were performed. The rats brain were removed to be subjected to the analysis of conventional HE staining, GAFP, S-100 immunohistochemical staining. At the same time, C6-Luc cells were inoculated in nude mice to detect the growth o