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>
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
Similar
MEDLINE
...
LILACS
LIS