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  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

  基金项目:高等学校博士学科点专项科研基金(20101106110031) 作者简介:全毅(1985-),男,硕士,主要研究方向:脑肿瘤治疗 通信联系人:高钟镐(1963-),男,研究员,主要研究方向:肿瘤靶向制剂和纳米给药系统.

  (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 30 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 of glioma. Tumor growth was detected 7 d after inoculation. Body weight and behavior of rats both had changes. The results of pathological examination showed that tumor cells can be seen along the nerve fibers to the surrounding invasion, and multiple hemorrhages were found in tumor tissues. Immunohistochemical staining showed that the expressions of glial fibrillary acidic protein, GFAP, and nerve fibers strongly positive S-100 protein and tumor cell aggregation were found, and blood-brain barrier was severely damaged along with the invasive growth of the tumor tissue to the opposite side. Most rats died within 30d. The C6/Wistar brain glioma animal model based on in vivo bioluminescence imaging was successfully constructed stably. It can be applied to evaluate anticancer drugs or gene therapy for pre-clinical study.

  Key words: glioma ; animal model ; bioluminescence imaging ; luciferase;Wistar rat

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