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目的:如何证实骨髓间充质干细胞(BMSCs)是胶质瘤基因治疗中最好的药物载体?将增强型绿色荧光蛋白(EGFP)标记的大鼠骨髓间充质干细胞移植入大鼠C6胶质瘤模型脑内,观察骨髓间充质干细胞在肿瘤内的迁徙与定位。方法:贴壁法培养大鼠骨髓细胞获取纯化的BMSCs。慢病毒介导EGFP转染BMSCs,于荧光显微镜下观察EGFP的表达,并行流式细胞仪检测EGFP阳性转染率。利用立体定向仪将培养好的C6细胞注入大鼠脑内,建立大鼠脑内胶质瘤模型。将标记EGFP的BMSCs利用微量注射器注入模型鼠脑内;移植后第1,7天处死大鼠,用荧光显微镜观察BMSCs在肿瘤内的迁移分布。结果:实验成功建立了大鼠脑内胶质瘤模型。以EGFP标记的BMSCs在模型鼠脑内主动迁移分布于肿瘤内部及肿瘤与正常脑组织交界侧。结论:骨髓间充质干细胞可以作为肿瘤基因治疗的良好载体。
OBJECTIVE: How to confirm that bone marrow mesenchymal stem cells (BMSCs) are the best drug carriers for gene therapy of glioma? Transplantation of enhanced green fluorescent protein (EGFP) labeled rat bone marrow mesenchymal stem cells into C6 glia Tumor model brain, bone marrow mesenchymal stem cells in the migration and positioning. Methods: Rat bone marrow cells were cultured with adherent method to obtain purified BMSCs. Lentivirus-mediated transfection of EGFP into BMSCs was used to observe the expression of EGFP under a fluorescence microscope. The transfection efficiency of EGFP was detected by flow cytometry. The cultured C6 cells were injected into the rat brain using a stereotactic instrument to establish a glioma model in the rat brain. BMSCs labeled with EGFP were injected into the brain of model mice with a microinjector. Rats were sacrificed on the 1st and 7th days after transplantation, and the distribution of BMSCs in the tumor was observed under a fluorescence microscope. Results: The rat brain glioma model was established successfully. EGFP-labeled BMSCs actively migrated in the brain of model mice and distributed inside the tumor and at the junction of tumor and normal brain tissue. Conclusion: BMSCs can be used as a good vector for gene therapy of tumors.