脑源性神经营养因子基因转染培养肌母细胞的表达

Expression of brain-derived neurotrophic factor in myoblasts transfected by PSVCEP BDNF-CAT with lipofect AMINE

目的 了解含脑源性神经营养因子 (BDNF)基因的表达质粒PSVCEPBDNF CAT在培养肌母细胞中的表达情况及LipofectAMINE阳离子脂质体介导的转染效率。 方法 按每 2 μg表达质粒PSVCEPBDNF CAT由 2 0 μlLipofectAMINE包裹后转染至细胞浓度为 2× 1 0 8个 /L的3 .5cm培养皿培养的肌母细胞中 ,连续观察细胞形态 ,48h后用免疫细胞化学和免疫电镜的方法来检测其表达情况。结果 培养肌母细胞在转染 6h吞噬脂质体最显著 ,转染 48h后 ,40 %的肌母细胞胞质内有BDNF基因表达 ,免疫电镜结果证实表达在细胞质近胞膜处。结论 含BDNF微基因的表达质粒PSVCEPBDNF CAT可以在培养肌母细胞中表达 ,脂质体转染的方法简便、有效。

Objective To study the expression of brain derived neurotrophic factor (BDNF) in myoblasts transfected by PSVCEP BDNF CAT and the transfecting efficiency of Lipofect AMINE.Methods 2?μg PSVCEP BDNF CAT was delivered into cultured myoblasts in 3.5 cm culture plate with cell concentration of 2×10 8/L by 20 μl Lipofect AMINE. The expression of BDNF in the transfected myoblasts was detected using immunocytochemistry and immuno electron microscopy.Results Lipofect AMINE was engulfed by myoblasts after 6 h remarkably; 48 h later, about 40% myoblasts could be detected with the expression of BDNF by immuno electron microscope and using immunocytochemistry.Conclusion PSVCEP BDNF CAT can express BDNF in the cultured myoblasts, and Lipofect AMINE is a convenient vector for gene transfection with simplicity and efficacy.