大鼠胚胎神经干细胞的培养及体外基因转染研究

The Cuiture of Nerve Trunk Cellc of Rats Embryo and in Gene Transfection

目的探讨大鼠胚胎神经干细胞的培养方法和经脂质体介导转染VEGF121基因的大鼠胚胎神经干细胞体外基因表达特点。方法选取孕14天SD大鼠胚胎海马,机械消化后在神经干细胞培养液中悬浮培养,并传代。脂质体介导法将VEGF121基因转染到大鼠神经干细胞中,经RT-PCR检测转基因神经干细胞体外基因表达时间窗;免疫荧光染色观察神经干细胞及其分化细胞的基因表达情况。结果培养获得高纯度神经干细胞、转基因神经干细胞及其分化的子代细胞均有VEGF121的表达并持续两周左右。结论经脂质体介导转染VEGF121基因的大鼠胚胎神经干细胞在体外良好表达基因产物。

Objective To culture embryonic neural stem cells（NSCs） of rats and detect gene expression of NSCs which is gene transferred of Vascular endothelial growth factor（VEGF） by lipofectamine technique. Methods Hippocampus of E14 rats were isolated and culured in NSC culture medium for over 6 passage. VEGF121 gene was transferred to rat NSCs by using lipofectamine technique. Gene expression of transfected cells and their progeny were detected by RT-PCR and immunofluorescent staining in vitro, Results NSCs was cultured successfully and VEGF-transfected NSCs could continuously express gene products during the first 2 weeks. Both transfected NSCs and their progeny expressed VEGF gene products, which was demonstrated by fluorescence study. Conclusion NSCs which are transfected VEGF121 gene by lipofectamine can express gene products efficiently in vitro.