摘要
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。
The stem cells of skeletal muscle from SD rats aged 2-4days were grown in a cultural plate for 24-48 hours. Using lipid-mediated DNA transfectio technique,the primary muscle cells were trans fected in vitro with plasmid which contained the promoter of Rous virus and β-galactosidase(LacZ)gene(pRSVLacZ).After 3-5 days,the cutured muscle cells were harvested and grafted into the caudate nucleus of adult SD brain. The implanted amount was 15ul containing 10￣6 cells for each case. The sections of brain grafted were checked after different postoperative survivals from 2 weeks to 24 weeks by X-gal histochemical staining for showing the activities of β-galactosidase and actin immunocytochemical staining for showing survival of the muscle cells. The results showed that approximately 40%of cultured muscle cell were transfected with pRSV LacZ in vitro and the plasmid expression in well survived muscle cells was stable for 24 weeks in vivo.These results suggested that the cultured cells should be useful as a vehicle for transgene expression in the brain and the method of using cacium liposome was desirable technique for gene transfer.
出处
《解剖学报》
CAS
CSCD
北大核心
1996年第2期140-143,共4页
Acta Anatomica Sinica
基金
国家自然科学基金
北京市自然科学基金
北京市科委854670100资助
