论文部分内容阅读
将新生２～３ｄ的ＳＤ大鼠骨骼肌细胞以１×１０￣６密度在培养皿中培养２４～４８ｈ，用阳离子脂质体介导技术将带有β－半乳糖苷酶基因的质粒ＤＮＡ导入这些培养肌细胞。３～５ｄ后，将经转基因的肌细胞按每１５μｌ含１×１０￣６个细胞数移植到ＳＤ大鼠脑内一侧纹状体中。术后动物分组存活２周、４周、８周、１２周、２０周和２４周后，取脑切片，用Ｘ－ｇａｌ组织化学法显示β－半乳糖苷酶活性。结果表明，采用脂质体对体外培养肌细胞的基因转移率可达４０％。将这些细胞植入动物脑内后从２周到２４周都有外源基因表达。本研究结果说明，骨骼肌细胞是表达外源性基因的良好载体细胞，而阳离子脂质体是转基因的有效介导物，它们为对某些神经系统病的基因治疗提供了可行途径。
Newborn 2 ~ 3d SD rat skeletal muscle cells were cultured in Petri dishes at a density of 1 × 10 ~ 6 for 24 ~ 48h. Plasmid DNA with β-galactosidase gene was introduced by cationic liposome-mediated technique These train muscle cells. After 3 to 5 days, the transgenic myocytes were transplanted into the striatum of one side of the brain of SD rats with 1 × 10 ~ 6 cells per 15μl. After 2 weeks, 4 weeks, 8 weeks, 12 weeks, 20 weeks and 24 weeks after operation, the animals were sacrificed and the brain slices were taken. The β-galactosidase activity was observed by X-gal histochemical method. The results showed that the gene transfer rate of liposomes in vitro muscle cells up to 40%. Exogenous gene expression occurs from 2 weeks to 24 weeks after these cells are implanted into the brain of animals. Our results demonstrate that skeletal muscle cells are good vector cells for expressing exogenous genes, while cationic liposomes are effective mediators of transgenes and provide a viable approach to the gene therapy of certain neurological diseases.