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绿色荧光蛋白转基因雄鼠肝细胞移植重建雌鼠的造血功能研究
  • ISSN号:1005-8982
  • 期刊名称:中国现代医学杂志
  • 时间:2013.7.20
  • 页码:12-15
  • 分类:R392.4[医药卫生—免疫学;医药卫生—基础医学]
  • 作者机构:[1]解放军昆明总医院干细胞工程实验室,云南昆明650032
  • 相关基金:国家自然科学基金(No:31172170)); 中国博士后科学基金(No:201104748)
  • 相关项目:Ⅱ型糖尿病的猕猴模型构建及重编程干细胞的效应机制
中文摘要:

目的用荧光雄鼠的肝细胞回输半致死量射线照射的C57BL雌鼠,观察肝细胞是否有重建造血功能的作用。方法用荧光雄鼠的肝细胞回输给半致死量射线照射的C57BL雌鼠,在移植后18、39和53 d剪鼠尾,采血并用CD4-PE和CD8-PE标记,再用氯化铵溶血,上流式细胞仪检测。移植87 d后杀鼠取骨髓用荧光原位杂交检测Y染色体阳性细胞的百分比,并取外周血、脾细胞、肝细胞和骨髓细胞检测绿色荧光蛋白(GFP)阳性细胞的百分比。结果回输肝细胞的雌鼠外周血中检测到GFP阳性细胞,同时检测到一部分GFP阳性细胞转化成了CD4+或CD8+T细胞,外周血中GFP阳性细胞的比例与荧光原位杂交分析的骨髓中的Y染色体阳性比例呈正相关。同时在鼠的脾细胞、肝细胞和骨髓细胞中都检测到了GFP阳性细胞。而脾细胞中检测到的GFP阳性细胞百分比高于其他细胞,相比有统计学意义(P=0.003)。结论成年鼠肝细胞含有一定量的造血干细胞,在移植鼠体内存活并分布在多脏器,重建了雌鼠的造血功能。

英文摘要:

【Objective】 Whether liver cells have hematopoietic reconstruction function was investigated by infusion of liver cells of fluorescent male mice into half-lethally irradiated C57BL female mice.【Methods】 Liver cells from fluorescent male mice were transfused back to half-lethally irradiated C57BL female mice.At 18,39,53 days after transplantation,mice were cut tails to collect blood samples,and labeled with CD4-PE and CD8-PE,and then bloods were hemolyzed with ammonium chloride.Then cells were detected by flow cytometry.87 days after transplantation,the mice were sacrificed to collect bone marrow to be detected Y chromosome positive percentage by fluorescence in situ hybridization.GFP positive cells rate were detected in peripheral blood,spleen cells,liver cells and bone marrow cells of the sacrificed mice.【Results】 GFP-positive cells were detected in the peripheral blood of female mice infused with liver cells.Some GFP-positive cells converted into CD4+ or CD8+ T cells.The percentage of GFP-positive cells was positively correlated with the proportion of Y chromosome cells in bone marrow.GFP positive cells can also be detected in peripheral blood,spleen cells,liver cells and bone marrow cells of the sacrificed mice.The percentage of GFP-positive cells detected in the spleen cells was higher than other cells with a statistical significance(P =0.003).【Conclusion】 The liver cells from adult mice contain a certain amount of hematopoietic stem cells.These cells survived in female mice and distributed in various organs,and reconstructed hematopoietic function.

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期刊信息
  • 《中国现代医学杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中华人民共和国教育部
  • 主办单位:中南大学 中南大学 中华人民共和国卫生部肝胆肠外科研究中心
  • 主编:陈子华
  • 地址:湖南省长沙市湘雅路87号
  • 邮编:410008
  • 邮箱:cjmm@2118.cn
  • 电话:0731-4327993 4803066
  • 国际标准刊号:ISSN:1005-8982
  • 国内统一刊号:ISSN:43-1225/R
  • 邮发代号:42-143
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),波兰哥白尼索引,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:59532