Adeno-associated virus type 2 recombinant green fluorescent protein transfection of cultured neural stem cells in vitro.docVIP

 PAGE \* MERGEFORMAT 11 Adeno-associated virus type 2 recombinant green fluorescent protein transfection of cultured neural stem cells in vitro [Abstract] Objective To study the two recombinant adeno-associated virus (recombinant adeno-associated virus 2, rAAV-2) vectors can effectively transfected NSCs. Methods containing the enhanced green fluorescent protein (enhanced green fluorescent protein, EGFP) reporter gene in rAAV-2 (rAAV-2/EGFP), with multiplicity of infection (multiplicity of infection, MOI), respectively 1 * 104,1 * 105, 1 * 106 in vitro transfection of isolated and cultured rat fetal NSCs, were observed under a fluorescence microscope, EGFP expression; Simultaneous Detection of the daughter cell viability, multiplication, differentiation to assess the situation on their survival, proliferation and differentiation of ; as stable EGFP expression by flow cytometry rAAV-2/EGFP transfection efficiency of the NSCs. The results 10 days after transfection the transfected group cloned the ball can be observed in NSCs issued a green fluorescence, initial fluorescence intensity varies with the increase of the value of enhanced MOI; each transfection group with the time of the fluorescence intensity gradually increased to 21 days after transfection and reached the peak and to maintain, this time by flow cytometry rAAV-2 Dui NSCs transfection efficiency: MOI for 1 * 104,1 * 105,1 * 106 when the transfection rate was 25.68% , 50.60%, 54.72%, fluorescence index (FI), respectively 4.08,12.72,14.06; transfection group and non-transfection group cell culture 7,14,21 days of their cell viability, multiplication, differentiation was no significant difference. Conclusion rAAV-2 can effectively transfected NSCs, mediated target gene expression can be highly efficient. [Keywords:] neural stem cells; 2-type adeno-associated virus; Transfection; multiplicity of infection; green fluorescent protein; fluorescence index; cell viability [Abstract] Objective To investigate