| GDNF慢病毒载体感染食蟹猴骨髓间充质干细胞 |
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| 引用本文： | 王玉兰,徐艳玲,任振华. GDNF慢病毒载体感染食蟹猴骨髓间充质干细胞[J]. 细胞生物学杂志, 2011, 0(1) |
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| 作者姓名： | 王玉兰 徐艳玲 任振华 |
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| 作者单位： | 首都医科大学宣武医院中心实验室;首都医科大学宣武医院细胞生物室;安徽医科大学人体解剖学教研室; |
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| 基金项目： | 北京市科委科技计划(No.D07050701350703)资助项目~~ |
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| 摘 要： | 骨髓间充质干细胞(mesenchymal stem cells,MSCs)是基因工程和细胞治疗的种子细胞之一,本研究利用含胶质源性神经营养因子(glial cell derived neurotrophic factor,GDNF)基因的慢病毒载体感染成年食蟹猴MSCs,探讨转染后GDNF在MSCs中的体外表达水平及其影响因素。首先,通过密度梯度离心法分离食蟹猴骨髓单核细胞(marrow mononuclear cells,MNCs),体外培养食蟹猴MSCs。同时构建表达GDNF的慢病毒载体,并感染食蟹猴MSCs,分别利用酶联免疫吸附(ELISA)方法和Real-time PCR方法,测定感染不同拷贝数病毒和不同转染组细胞GDNF的蛋白分泌水平和基因表达水平。实验结果显示,表达GDNF基因的慢病毒载体成功转染成年食蟹MSCs,体外培养的MSCs持续表达分泌GDNF。感染慢病毒的拷贝数可以影响GDNF分泌水平,相同条件下感染拷贝数越高,GDNF分泌量越多,其基因表达水平越高。
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| 关 键 词： | 食蟹猴 骨髓间充质干细胞 胶质源性神经营养因子 慢病毒载体 感染 |
GDNF Lentivirus Infection of Cynomolgus Monkey Bone Marrow Mesenchymal Stem Cells |
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| Yu-Lan Wang,Yan-Ling Xu,Zhen-Hua Ren. GDNF Lentivirus Infection of Cynomolgus Monkey Bone Marrow Mesenchymal Stem Cells[J]. Chinese Journal of Cell Biology, 2011, 0(1) |
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| Authors: | Yu-Lan Wang Yan-Ling Xu Zhen-Hua Ren |
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| Affiliation: | Yu-Lan Wang~1,Yan-Ling Xu~2,Zhen-Hua Ren~(2,3*) (1 Central Laboratory,Xuanwu Hospital,Capital Medical University,Beijing 100053,China,2 Cell Biology Laboratory,3 Department of Anatomy,Anhui Medical University,Hefei 230032,China) |
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| Abstract: | Mesenchymal stem cells(MSCs) have received considerable attention for various therapeutic approaches in recent years.MSCs are also easy to genetically modify to express therapeutic genes using lentiviral vectors.In this study,we transfected cynomolgus monkey mesenchymal stem cells(MSCs) using lentiviral vectors encoding glial cell line derived neurotrophic factor(GDNF) to study its expression level in vitro.First,Lentiviral vectors encoding GDNF were packaged by 293T cells through three plasmids transient c... |
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| Keywords: | cynomolgus monkey mesenchymal stem cells glial cell derived neurotrophic factor lentiviral vectors transduction |
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