| Biomedicine |
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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 |
| Yong FU, Shen-qing WANG, Ying-peng LIU, Guo-peng WANG, Jian-ting WANG, Shu-sheng GONG |
| Department of Otorhinolaryngology, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China; Department of Otorhinolaryngology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China; Department of Surgery-Otolaryngology, University of Kentucky Medical Center, Lexington, Kentucky 40536, USA; Department of Surgery-Otolaryngology, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China |
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Abstract 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. Conclusion: 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.
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Received: 22 August 2007
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