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Gene transfer into primary cultures of fetal neural stem cells by a recombinant adenovirus carrying the gene for green fluorescent protein / 浙江大学学报(英文版)(B辑:生物医学和生物技术)
Journal of Zhejiang University. Science. B ; (12): 299-305, 2008.
Article in English | WPRIM | ID: wpr-359429
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the transduction efficiency of a recombinant adenovirus carrying the gene for green fluorescent protein (Ad-GFP) into the primary cultures of fetal neural stem cells (NSCs) by the expression of GFP.</p><p><b>METHODS</b>The Ad-GFP was constructed by homologous recombination in bacteria with the AdEasy system; NSCs were isolated from rat fetal hippocampus and cultured as neurosphere suspensions. After infection with the recombinant Ad-GFP, NSCs were examined with a fluorescent microscopy and a flow cytometry for their expression of GFP.</p><p><b>RESULTS</b>After the viral infection, flow cytometry analysis revealed that the percentage of GFP-positive cells was as high as 97.05%. The infected NSCs sustained the GFP expression for above 4 weeks. After differentiated into astrocytes or neurons, they continued to express GFP efficiently.</p><p><b>CONCLUSION</b>We have successfully constructed a viral vector Ad-GFP that can efficiently infect the primary NSCs. The reporter gene was showed fully and sustained expression in the infected cells as well as their differentiated progenies.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Stem Cells / Recombinant Proteins / Adenoviridae / Chemistry / Gene Transfer Techniques / Cell Biology / Green Fluorescent Proteins / Electrophoresis, Agar Gel / Escherichia coli / Genetic Vectors Limits: Animals / Humans Language: English Journal: Journal of Zhejiang University. Science. B Year: 2008 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Stem Cells / Recombinant Proteins / Adenoviridae / Chemistry / Gene Transfer Techniques / Cell Biology / Green Fluorescent Proteins / Electrophoresis, Agar Gel / Escherichia coli / Genetic Vectors Limits: Animals / Humans Language: English Journal: Journal of Zhejiang University. Science. B Year: 2008 Type: Article