| 小鼠脑皮质神经干细胞体外培养及miRNAs影响其分化能力 |
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| 引用本文： | 赵祥宇,舒鹏程,张靖,阴彬,彭小忠.小鼠脑皮质神经干细胞体外培养及miRNAs影响其分化能力[J].基础医学与临床,2011,31(6). |
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| 作者姓名： | 赵祥宇 舒鹏程 张靖 阴彬 彭小忠 |
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| 作者单位： | 中国医学科学院,基础医学研究所,北京协和医学院,基础学院,医学分子生物学国家重点实验室,北京,100005 |
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| 基金项目： | 国家自然科学基金(31071203) |
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| 摘 要： | 目的系统建立小鼠脑皮质神经干细胞(NSCs)体外培养的技术平台,并探索一组microRNA对NSCs分化能力的影响。方法胎鼠(E14)脑皮质分离NSCs进行体外培养;免疫荧光法检测NSCs向神经元和星型胶质细胞的自然分化;Real-time定量PCR筛选一组分化前后表达水平有显著性差异的microRNA;应用新一代脂质体高效转染NSCs,将该组microRNA或其抑制物导入NSCs;Western blot检测导入的RNA序列对NSCs分化能力的影响。结果基于小鼠NSCs体外培养模型,筛选出一组NSCs分化前后表达水平存在显著性差异的microRNA(miR-124,137,128)。Western blot结果显示miR-124的反义链能够明显下调β-tubulinⅢ的表达(P<0.01),过表达miR-137和miR-128后β-tubulinⅢ的表达量有所升高。结论脂质体转染技术能够将miRNAs高效导入NSCs中;microRNA-124,137,128能够影响NSCs的分化。
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| 关 键 词： | 神经干细胞 神经元 miRNA 脂质体转染 分化 |
Culture of mouse cortex neural stem cells and miRNAs' influence on NSCs' differentiation ability |
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| ZHAO Xiang-yu,SHU Peng-cheng,ZHANG Jing,YIN Bin,PENG Xiao-zhong.Culture of mouse cortex neural stem cells and miRNAs' influence on NSCs' differentiation ability[J].Basic Medical Sciences and Clinics,2011,31(6). |
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| Authors: | ZHAO Xiang-yu SHU Peng-cheng ZHANG Jing YIN Bin PENG Xiao-zhong |
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| Institution: | ZHAO Xiang-yu,SHU Peng-cheng,ZHANG Jing,YIN Bin,PENG Xiao-zhong(National Laboratory of Medical Molecular Biology,Institute of Basic Medical Sciences,CAMS & PUMC,Beijing 100005,China) |
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| Abstract: | Objective To establish the mouse cortex neural stem cells(NSCs) culture technology platform for exploring miRNA' influence on neural stem cells' differentiation ability.Methods The NSCs were prepared after isolation from the E14 embryonic mouse cortex and cultured in proliferation medium,the cells were then cultured in the differentiation medium with the immunostaining method to detect whether they can differentiate into neurons and astrocytes normally.Meanwhile Realtime-PCR was used to detect the expressio... |
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| Keywords: | neural stem cells neurons miRNA transfection differentiation |
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