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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2018, Vol. 38 Issue (6): 58-62    DOI: 10.13523/j.cb.20180608
技术与方法     
内源CD133 +细胞示踪小鼠模型的制备和鉴定 *
韩明明,罗玉萍()
南昌大学生命科学学院 南昌 330031
Establishment and Identification of Endogenous CD133 + Cell Tracer Mouse Model
Ming-ming HAN,Yu-ping LUO()
School of Life Science,Nanchang University,Nanchang 330031,China
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摘要：

目的:为了制备可追踪CD133阳性神经干细胞分化谱系的小鼠模型。方法:将两种C57B16背景的转基因小鼠CD133-Cre-ERT2和Rosa26-CAG-LSL-ZsGreen杂交,获得CD133-CreER;CAG-ZsGreen小鼠模型。结果:免疫组化和激光扫描共焦成像分析表明,经Tamoxifen作用后,该杂交小鼠在侧脑室SVZ区、第三脑室和第四脑室的室管膜区域均表达绿色荧光蛋白ZsGreen,且这些区域的绿色荧光与CD133+红色荧光重合。 结论:在室管膜区CD133+是静息态神经干细胞的标志,因此,通过分析CD133-CreER;CAG-ZsGreen小鼠中的 ZsGreen阳性细胞可追踪神经干细胞的细胞分化谱系。成功制备了内源CD133+细胞示踪小鼠模型,为探讨大脑中CD133+神经干细胞的激活、增殖、迁移和分化提供了帮助。
关键词： CD133;  神经干细胞;  转基因小鼠    
Abstract:

Objective:To establish a mouse model that can trace the differentiation of CD133 positive neural stem cells.Methods:The CD133-Cre-ERT2 transgenic mice were mated with Rosa26-CAG-LSL-ZsGreen transgenic mice to produce CD133-CreER;CAG-ZsGreen double transgenic in C57B16 mice.Result:The immunohistochemistry and laser scanning confocal imaging analysis showed that Tamoxifen-Inducible Cre/loxP recombination, as depicted by green cells, existed in lateral ventricle SVZ, the ventricular zone of third ventricle and the fourth ventricle in adult mice, and the green fluorescence overlaps with CD133+ red fluorescence in these regions.Conclusion:CD133+ is the sign of the resting nerve stem cell in the ventricle membrane, so it can trace the cell differentiation lineages of neural stem cells by analyzing ZsGreen positive cells in CD133-CreER;CAG-ZsGreen mice. The mouse model for in vivo lineage tracing of endogenous CD133+ cell was established successfully, which will be helpful to explore the activation, proliferation, migration and differentiation of CD133+ neural stem cells in the brain.

Key words: CD133    Neural stem cells    Transgenic mouse
收稿日期: 2017-12-24 出版日期: 2018-07-06
ZTFLH:  Q78  
基金资助: * 国家自然科学基金(31660324);江西省自然科学基金重大项目资助项目(2015ACB20008)
通讯作者: 罗玉萍     E-mail: luoyuping@163.com
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引用本文:

韩明明,罗玉萍. 内源CD133 +细胞示踪小鼠模型的制备和鉴定 *[J]. 中国生物工程杂志, 2018, 38(6): 58-62.

Ming-ming HAN,Yu-ping LUO. Establishment and Identification of Endogenous CD133 + Cell Tracer Mouse Model. China Biotechnology, 2018, 38(6): 58-62.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20180608    或    https://manu60.magtech.com.cn/biotech/CN/Y2018/V38/I6/58

图1  CD133-Cre-ERT2和Rosa26-CAG-LSL-ZsGreen转基因小鼠的PCR鉴定结果
图2  CD133阳性细胞示踪小鼠制备示意图
图3  免疫组化染色显示ZsGreen和CD133+神经干细胞在小鼠室管膜区的分布
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