Co-expression of green hypoxia inducible factor-1αand green fluorescent protein in neural stem cells

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Co-expression of green hypoxia inducible factor-1αand green fluorescent protein in neural stem cells

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- Co-expression of green hypoxia inducible factor-1αand green fluorescent protein in neural stem cells
- Journal of Chongqing Medical University Issue 11, Pages: 1138-1142(2007)
- 作者机构：
  
  1. 重庆医科大学附属第二医院神经内科
  2. 重庆医科大学附属第二医院神经内科,重庆,400010
- 作者简介：
- 基金信息：
- DOI：
  CLC： R363
- Published：2007，
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[1]吴万福,陈秀,胡长林.缺氧诱导因子-1α与绿色荧光蛋白在神经干细胞中的共表达[J].重庆医科大学学报,2007(11):1138-1142.

WU Wanfu, et al. Co-expression of green hypoxia inducible factor-1αand green fluorescent protein in neural stem cells. [J]. Journal of Chongqing Medical University (11):1138-1142(2007)
[1]吴万福,陈秀,胡长林.缺氧诱导因子-1α与绿色荧光蛋白在神经干细胞中的共表达[J].重庆医科大学学报,2007(11):1138-1142. DOI：

WU Wanfu, et al. Co-expression of green hypoxia inducible factor-1αand green fluorescent protein in neural stem cells. [J]. Journal of Chongqing Medical University (11):1138-1142(2007) DOI：

摘要

目的:以携带缺氧诱导因子-1α（Hypoxia-inducible factor-1α

HIF-1α）及绿色荧光蛋白（Green fluorescent protein

GFP）基因的腺病毒转染大鼠神经干细胞（Neuralstemcells

NSCs）

观察HIF-1α和GFP在NSCs中的表达以及对NSCs生物学特性的影响

为HIF-1α基因转染的NSCs移植治疗缺血性脑血管病提供物质基础。方法:利用携带HIF-1α和GFP基因的重组腺病毒转染NSCs

相差显微镜下观察NSCs的形态及在荧光显微镜下观察NSCs及其诱导分化后的GFP表达情况;观察HIF-1α基因在NSCs中的表达;并检测NSCs的细胞活性。应用免疫荧光技术检测神经干细胞及其诱导后分化细胞的特异性蛋白巢蛋白（Nestin）、神经微丝蛋白（Neurofilamentprotein）和胶质纤维酸性蛋白（GFAP）。结果:重组腺病毒转染NSCs后外源性基因及绿色荧光蛋白有持续稳定表达;转染后的NSCs的形态学、细胞活性和分化能力等与未转染的NSCs无明显差异（P>0.05）;检测神经元标志物神经微丝蛋白和胶质细胞标志物胶质纤维酸性蛋白

证实转染后的NSCs仍然具有多向分化潜能。结论:NSCs转染携带HIF-1α和GFP基因的重组腺病毒后能够持续、稳定地表达HIF-1α和GFP

生物学特征正常。

Abstract

Objective:To provide material foundation for the transplantation of neural stem cells(NSCs)in the treatment of cerebral ischemia

we transfected NSCs using recombinant adenovirus with HIF-1α and GFP gene

and observed the expression of HIF-1α and GFP

and their effects on bionomics of the NSCs.Methods:NSCs were transfected with the recombinant adenovirus

and then the expression of HIF-1α and GFP in the NSCs was observed.NSCs were detected under phase contrast microscope and fluorescent microscope.The special protein such as nestin

neurofilment protein(NFP)

glial fibriliary acidic protein(GFAP)were detected with immunofluorenscence technique.Results:The exogenous gene could be expressed steadily in NSCs after transfection.The morphological features of the untransfected NSCs were similar to those of the transfected

and there were no significant differences between the transfected NSCs and the untransfected in the cytoactivity and differentiation(P>0.05).The transfected NSCs were confirmed to have the latent ability of multi-directional differentiation.Conclusion:NSCs can express HIF-1α and GFP steadily after transfected with the recombinant adenovirus and the bionomics is normal.

关键词

神经干细胞缺氧诱导因子-1α绿色荧光蛋白重组腺病毒

Keywords

Neural stem cells(NSCs)Hypoxia inducible factor-1α(HIF-1α)green fluorescent proteinrecombinant adenovirus

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