重组rAAV2/eGFP、rAAV2/NGF病毒转染神经干细胞的研究

目的　观察重组rAAV2 /eGFP(含报告基因 )和rAAV2 /NGF(含目的基因 )病毒转染神经干细胞的转染效率 ;探讨神经干细胞治疗神经系统疾病的新途径。方法　分离、培养和鉴定神经干细胞后 ,用重组rAAV2 /eGFP和rAAV2 /NGF病毒转染神经干细胞 ,用荧光显微镜和免疫细胞化学检测GFP和NGF在体外的表达。结果　GFP和NGF于 2 4h后在体外开始表达 ;5d后表达效率高达 89% ,并可持续稳定表达 35d以上 ;MOI(v g/cell)不同 ,表达效率不同。 结论　重组rAAV2 /eGFP和rAAV2 /NGF病毒可转染神经干细胞 ,神经干细胞可稳定表达GFP和NGF ,可用于基因治疗神经系统疾病。

Study on Transfecting of Neural Stem Cells by Recombined rAAV2/eGFP and rAAV2/NGF Virus

Objective To observe the rate of transfec ti on of recombined rAAV2/eGFP(containing reporting genes) and rAAV2/NGF(containing targeting genes) virus into neural stem cells to explore the ways of neural ste m cells treating nervous system diseases. Method Neural stem cells were transfected by recombined rAAV2/eGFP and rAAV2/NGF virus after separa tion,culture and characterization,then we detected the expression of GFP and NGF by fluorescence microscopy and immunocytochemistry in vitro. Results GFP and NGF began to express 24 hours after transfection in vitro;the r ate of expression rose to 89% 5 days later,and could last for more than 35 days ;the rate of expression of them was different with the different MOI(v.g./cell) . Condusion Recombined rAAV2/eGFP and rAAV2/NGF virus can be transfected into neural stem cells which could express GFP,NGF steadly and may play a important role in therapy of nervous system diseases.