间充质干细胞双标记光学成像的体内试验研究

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目的:验证双标记生物发光成像活体观测MSCs在肝癌裸鼠模型向肿瘤病灶的趋化作用的可行性。方法:应用fluorescence(荧光)与bioluminescence(生物发光)两种成像方法,对MSCs进行CM-Di I荧光标记及对人肝癌细胞Hep G2进行Fluc-慢病毒感染并由此建立裸鼠肝癌模型,构建双标记成像系统,应用精诺真小动物光学成像仪在裸鼠肝癌模型中观测间充质干细胞向肿瘤的趋化作用。结果:在鼠尾静脉注射标记MSCs细胞后21天荧光成像可见MSCs主要积聚于肿瘤病灶处及肝脏。生物发光成像后可监测到病灶处由luciferase标记肿瘤细胞(Hep G2)发出荧光;将荧光成像与生物发光成像所得图像经后处理融合后,可见证间充质干细胞像肿瘤病灶定向迁徙的生物过程。经肿瘤病理切片证实间充质干细胞成功迁徙至肿瘤病灶中。结论:应用间充质干细胞双标记光学成像系统实现MSCs在活体内对肿瘤的趋化过程进行观测是可行的。这种成像方法可作为下一步以MSCs为载体的肿瘤基因治疗的有效监测手段。 OBJECTIVE: To verify the feasibility of double labeled bioluminescence imaging to observe the chemotactic effect of MSCs in tumor-bearing hepatocellular carcinoma model in nude mice. Methods: Fluorescence and bioluminescence imaging were used to detect MSCs labeled with CM-Di I and Fluc-lentivirus infection of human hepatocellular carcinoma cell line Hep G2. Dual-labeling imaging system, the application of real minnow animal optical imager in nude mouse liver cancer model observed mesenchymal stem cell chemotaxis to the tumor. Results: After MSCs were injected into the caudal vein for 21 days, MSCs mainly accumulated in the lesion and the liver. After bioluminescence imaging, luciferase-labeled tumor cells (Hep G2) can be detected in the lesion. After fluorescence imaging and bioluminescence imaging, the images obtained by post-treatment fusion can be seen in the biological process of directional migration of mesenchymal stem cells . Confirmed by the tumor biopsy mesenchymal stem cells successfully migrate to the tumor lesions. Conclusion: It is feasible to use MSCs dual-labeled optical imaging system to observe the chemotactic process of MSCs in vivo. This imaging method can be used as the next step to MSCs as a carrier of cancer gene therapy effective monitoring means.
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