快速高效构建小鼠体内成像肿瘤模型的方法学平台  

A rapid and efficient method for constructing murine tumor models with in vivo imaging

作　　者：张胤晟[1] 林丹丹[1] 雷蕾[1] 胡博[1] 王仲娟[1] 刘海燕[1,2] 

机构地区：[1]苏州大学生物医学研究院肿瘤细胞与分子免疫学实验室,215123 [2]苏州大学第一附属医院江苏省血液研究中心,215006

出　　处：《免疫学杂志》2013年第4期346-350,共5页Immunological Journal

基　　金：国家自然科学基金重大研究计划培育项目(91029703)

摘　　要：目的建立小鼠肿瘤模型,并用体内成像的方法进行检测,降低传统方法中的系统误差与人为测量错误,提高模型的准确性与客观性。方法构建萤火虫素酶的慢病毒表达载体,体外包装能表达萤火虫素酶的慢病毒颗粒。用病毒颗粒转染目标细胞,筛选出稳定表达萤火虫素酶的目标细胞系。体外运用双荧光素酶分析系统进行质控检测,通过后建立小鼠肿瘤模型,最后使用活体成像的方法对小鼠肿瘤模型中肿瘤的大小进行定量分析,以评估小鼠肿瘤模型的恶化程度。结果成功构建含有萤火虫素酶基因的慢病毒质粒;包装能正确表达具有功能活性萤火虫素酶的慢病毒颗粒;使用病毒颗粒感染目标细胞并筛选出能表达萤火虫素酶的稳转细胞系;建立小鼠皮下肿瘤模型,并利用活体成像的方法对肿瘤大小进行定量分析。结论通过该种方法可以建立表达萤火虫素酶的稳转细胞系,利用体内成像的检测方法可以准确的对肿瘤模型的发生发展进行评估。In vivo imaging allows continuous and accurate measurement of tumor growth without sacrificing animals. However, the tumor cell lines expressing luciferase or fluorescence commercially available are extremely limited. By using a lentiviral vector expressing luciferase and yellow fluorescent protein （YFP） constructed previously by our laboratory, lentivirus was packaged and used for infecting target cell line hepa1-6. Then the infected cells were screened and selected for high YFP expression by flow sorting, as well as for stable and robust luciferase expression. The stable cell line was injected subcutaneously into C57/BL6 mice to generate tumor models with in vivo imaging. Luciferase could be stably expressed by the target ceils after lentiviral infection, and the high expressing cells could be sorted by YFP expression. Thus tumor model with in vivo imaging was successfully established using the infected cell line. In conclusion, this method reported here can be used to generate tumor models with in vivo imaging rapidly and efficiently.

关 键 词：小鼠肿瘤模型 慢病毒 活体成像 萤火虫素酶 

分 类 号：R73[医药卫生—肿瘤]