阳离子脂质体转染人类骨骼肌原代细胞的初步研究

探讨不同脂质体介导基因转染人类骨骼肌原代细胞的转染效率和基因的表达.将含有β-半乳糖苷酶LacZ结构基因的质粒,用三种不同的阳离子脂质体导入人类骨骼肌原代细胞中,通过X-Gal染色观察不同的转染效率.结果发现,Fugene 6转染效率最高,蓝染细胞达10%,其脂质体与DNA的最佳比例为3∶ 2.Fugene 6可有效地将外源基因导入骨骼肌原代细胞,而且外源基因可以长效高效地表达,有望用来作为基因治疗的载体.

Study on the transfection of primary human skeletal muscle cells with different cationic liposome

To study the transfection efficiency and the expression of transgene in primary human skeletal muscle cells with different liposome,the plasmid which contained LacZ gene was transferred into cells with three different cationic liposome.Transfection efficiency was determined by X-gal staining.Results showed that the transfection efficiency in Fugene 6 group was the highest than that in other groups(10%),the best ratio of Fugene 6 with DNA was 3: 2.Fugene 6 could effectively transfer the gene into primary human skeletal muscle cells.The transgene was still expressed after a longer period.Skeletal muscle cells can be used as the vector for gene therapy.