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作 者:林沪[1] 毛青[1] 王宇明[1] 蒋业贵[1] 邓国宏[1] 蒋黎[1]
机构地区:[1]第三军医大学西南医院感染病研究所,重庆400038
出 处:《中华肝脏病杂志》2006年第8期578-581,共4页Chinese Journal of Hepatology
基 金:国家自然科学基金(30271173)
摘 要:目的研究正常人L02肝细胞移植到具有正常免疫活性的大鼠肝内的存活情况,建立稳定的人鼠嵌合肝动物模型。方法SD大鼠出生前宫内腹腔注射L02肝细胞,诱导胎鼠对人L02肝细胞产生免疫耐受,出生2周后经脾移植Dit染色后的人L02肝细胞,建立人鼠嵌合肝动物模型。采用免疫荧光、免疫组织化学、DiI荧光示踪等方法,在不同时相分别检测人白蛋白和特异性人增殖细胞核抗原,在荧光显微镜下观察人L02肝细胞在鼠肝内的分布。结果于移植后1、2、4、6、8、10周在荧光显微镜下观察到人L02肝细胞在鼠肝内的动态分布;移植后2、4,6、8周大鼠均检测出人白蛋白;移植后2、4、6周检测出特异性人增殖细胞核抗原。结论移植的人L02肝细胞在具有正常免疫活性的免疫耐受鼠体内能够存活、增殖,并产生人白蛋白。Objective To investigate whether human L02 hepatocytes could survive after implanting them into normal, immunocompetent rats. Methods Human L02 hepatocytes were injected through the uterine walls into the intraperitoneal cavities of fetal Sprague-Dawley rats to induce immune tolerance to human L02 hepatocytes. Human L02 hepatocytes stained with DiI were implanted into the spleens of the 2-week old rats. Immuno-fluorescent staining, SP immunohistochemistry, and DiI staining were used to detect human albumin and specific proliferating cell nuclear antigen (PCNA) in the rat livers. The distribution of human L02 hepatocytes was observed under the fluorescent microscope. Results Dynamic distribution of human L02 hepatocytes in the rat livers was observed from the 1st to the 10th week after the implantation. Human albumin was detected at 2, 4, 6 and 8 weeks, and at the 4th week it had the highest level. Specific human PCNA was detected in the rat livers from the 2nd to the 6th week after implantation. The PCNA positive cells were most aboundant at the 4th week. ConeIusion Human L02 hepatocytes can survive and proliferate for 10 weeks after implanting them into genetically normal immunocompetent rats.
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