文章摘要
荧光素酶标记小鼠结肠癌细胞建立活体成像移植瘤模型*
Establishment of a live imaging immunotherapy xenograft model by luciferase-labeled mouse colon cancer cells *
投稿时间:2018-12-18  修订日期:2019-01-03
DOI:
中文关键词: 荧光素酶  CT26细胞  移植瘤  活体成像  Balb/c鼠
英文关键词: Luciferase  CT26 cell  Xenografts  In vivo imaging  Balb/c mice
基金项目:国家自然科学基金面上项目(81472843)和上海市科学技术委员会上海市科研计划项目(14ZR1424700)
作者单位E-mail
罗华灶 复旦大学生命科学学院微生物感染与分子免疫学实验室 上海 16210700005@fudan.edu.cn 
李雪 复旦大学生命科学学院微生物感染与分子免疫学实验室 上海  
依含 复旦大学  
谢君 复旦大学生命科学学院微生物感染与分子免疫学实验室 上海  
朱乃硕 复旦大学生命科学学院微生物感染与分子免疫学实验室 上海 nzhu@fudan.edu.cn 
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中文摘要:
      目的:建立稳定表达荧光素酶的小鼠结肠癌细胞株并构建适用于小动物活体成像系统观察的Balb/c鼠皮下移植瘤模型。方法:通过基因重组的方式,将外源基因插入到逆转录病毒载体上,以工具细胞293T包毒后得到病毒,将携带荧光素酶基因的质粒转染到小鼠结肠癌细胞株CT26中,Puromycin筛选出稳定表达荧光素酶的单克隆细胞株。扩增后接种于Balb/c鼠,建立Balb/c鼠皮下移植瘤模型,通过活体动物成像系统监测肿瘤的生长过程。结果:获得了高水平稳定表达荧光素酶的结肠癌单克隆细胞株,该单克隆细胞株与母细胞系CT26具有相似的生长特性。将稳定表达荧光素酶的克隆接种于Balb/c鼠皮下可成瘤,小动物活体成像系统能准确监测肿瘤细胞在体内的生长过程。结论:成功建立了稳定表达荧光素酶的结肠癌单克隆细胞株。采用活体动物成像系统构建的Balb/c鼠皮下结肠癌移植瘤模型是拓展肿瘤体内生长、转移及治疗相关研究的理想模型。
英文摘要:
      Objective: To establish a mouse colon cancer cell line stably expressing luciferase and construct a Balb/c mouse subcutaneous transplantation tumor model suitable for small animal living imaging system. Methods: The foreign gene was inserted into the retroviral vector by gene recombination, and the virus was obtained by inoculating the tool cell 293T. The plasmid carrying the luciferase gene was transfected into mouse colon cancer cell line CT26, Puromycin. Monoclonal cell lines stably expressing luciferase were screened. After amplification, they were inoculated into Balb/c mice to establish a Balb/c mouse subcutaneous tumor model, and the tumor growth process was monitored by a living animal imaging system. Results: A colon cancer monoclonal cell strain stably expressing luciferase at a high level was obtained, which had similar growth characteristics to the parental cell line CT26. The clones stably expressing luciferase were inoculated into the skin of Balb/c mice to form tumors, and the small animal living imaging system can accurately monitor the growth process of tumor cells in vivo. Conclusion: A colon cancer monoclonal cell strain stably expressing luciferase was successfully established. The Balb/c mouse subcutaneous colon cancer xenograft model constructed by the living animal imaging system is an ideal model for expanding the research on growth, metastasis and treatment of tumors in vivo.
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