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[绿色荧光蛋白表达的人胶质瘤干细胞在裸鼠中的移植]

[Transplantation of human glioma stem cells in nude mice with green fluorescent protein expression].

作者信息

Wu Zi-cheng, Huang Qiang, Shao Yi-xiang, Xue Zhi-mou, Dong Jun, Diao Yi, Wang Ai-dong, Lan Qing

机构信息

Department of Neurosurgery, Second Affiliated Hospital of Suzhou University, Suzhou 215004, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2008 Aug 26;88(33):2317-20.

Abstract

OBJECTIVES

To investigate the possibility of transplantation of human glioma stem cells (HGSCs) in nude mice stably expressing green fluorescent protein (GFP) so as to clearly identify the incubated HGSCs from the host tissues.

METHODS

Transgenic C57BL/6J mice expressing GFP was crossed with nude mice of the line NC, then hairless male nude mice expressing GFP were crossed with hairy female pubescent mice to obtain nude mice with GFP expression the expression of GFP in the skin and organs of these nude mice were evaluated by naked eyes, and immunohistochemical and immunofluorescence assays. HGSCs were transplanted orthotopically into the caudate nuclei of nude mice expressing GFP. Immunohistochemistry was used to observe the transplanted tumor.

RESULTS

The structures rich in adipose tissue of the 8th generation nude mice were dark green and the other organs were light green. However, green fluorescence was emitted from all tissues under fluorescence microscopy. Confocal fluorescence microscopy showed that the tumor cells were stained red, distinguished from the host cells distinctly in the brains bearing tumor transplanted orthotopically.

CONCLUSION

Nude mice expressing GFP can be obtained by crossing the transgenic mice bearing naive immunity with nude mice. Orthotopic transplantation of HGSCs may be used in the investigation of tumor tissue reconstitution because of the easy identification between the transplantation tumor and host tissue.

摘要

目的

研究稳定表达绿色荧光蛋白(GFP)的人胶质瘤干细胞(HGSCs)在裸鼠体内移植的可能性,以便从宿主组织中清晰识别植入的HGSCs。

方法

将表达GFP的转基因C57BL/6J小鼠与NC品系裸鼠杂交,然后将表达GFP的无毛雄性裸鼠与有毛雌性青春期小鼠杂交,获得表达GFP的裸鼠,通过肉眼观察以及免疫组织化学和免疫荧光分析评估这些裸鼠皮肤和器官中GFP的表达情况。将HGSCs原位移植到表达GFP的裸鼠尾状核中。采用免疫组织化学方法观察移植瘤。

结果

第8代裸鼠富含脂肪组织的结构呈深绿色,其他器官呈浅绿色。然而,在荧光显微镜下所有组织均发出绿色荧光。共聚焦荧光显微镜显示肿瘤细胞被染成红色,在原位移植瘤的大脑中与宿主细胞明显区分开来。

结论

将具有天然免疫的转基因小鼠与裸鼠杂交可获得表达GFP的裸鼠。由于移植瘤与宿主组织易于鉴别,HGSCs原位移植可用于肿瘤组织重建的研究。

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