Gene transfer into primary cultures of fetal neural stem cells by a recombinant adenovirus carrying the gene for green fluorescent protein.
- Author:
Yong FU
1
;
Shen-qing WANG
;
Ying-peng LIU
;
Guo-peng WANG
;
Jian-ting WANG
;
Shu-sheng GONG
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviridae; metabolism; Animals; Electrophoresis, Agar Gel; Escherichia coli; metabolism; Gene Transfer Techniques; Genetic Vectors; Green Fluorescent Proteins; metabolism; Hippocampus; cytology; Humans; Models, Biological; Models, Genetic; Neurons; cytology; Rats; Recombinant Proteins; chemistry; Stem Cells; cytology
- From: Journal of Zhejiang University. Science. B 2008;9(4):299-305
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo 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.
METHODSThe 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.
RESULTSAfter 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.
CONCLUSIONWe 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.
