Gene transfer into primary cultures of fetal neural stem cells by a recombinant adenovirus carrying the gene for green fluorescent protein.
OBJECTIVE
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Fu
Yong
Y
Department of Otorhinolaryngology, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China. fyent@163.com
Wang
Shen-qing
SQ
Liu
Ying-peng
YP
Wang
Guo-peng
GP
Wang
Jian-ting
JT
Gong
Shu-sheng
SS
D000256:Adenoviridae / Q000378:metabolism*; D000818:Animals; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D008954:Models, Biological; D008957:Models, Genetic; D009474:Neurons / Q000166:cytology*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D013234:Stem Cells / Q000166:cytology*
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins