Lipofection of cultured mouse muscle cells: a direct comparison of Lipofectamine and DOSPER
Lipofection of cultured mouse muscle cells: a direct comparison of Lipofectamine and DOSPER
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DOI：
10.1038/sj.gt.3300604
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发表时间：
1998-04-01
期刊：
影响因子：
5.1
通讯作者：
Dickson, G
中科院分区：
文献类型：
--
作者：
Dodds, E;Dunckley, MG;Dickson, G
阳离子脂质-DNA复合物（lipoplexes）已被广泛用作基因转移载体，其避免了病毒载体的不良免疫原性和潜在的病毒血症。随着体内基因转移到骨骼肌的长期目标，我们描述了一个直接在体外比较两个市售的阳离子脂质制剂，脂质体和DOSPER。在C2 C12小鼠肌细胞系中进行转染的优化，然后在原代小鼠成肌细胞和C2 C12肌管中进行进一步研究。报告基因构建体表达E.大肠杆菌β-半乳糖苷酶或绿色荧光蛋白（GFP）分别通过组织化学染色或流式细胞术分析来评估转染效率。两种脂质制剂都能够促进C2 C12细胞中高效、可重复的基因转移，并成功地抑制原代小鼠成肌细胞培养物。然而，DOSPER表现出能够在牛和鼠来源的血清存在下转染细胞的重要优势。该特征允许在体外转染期间增加细胞存活，并且可能有利于直接体内基因转移功效。
Cationic lipid-DNA complexes (lipoplexes) have been widely used as gene transfer vectors which avoid the adverse immunogenicity and potential for viraemia of viral vectors. With the long-term aim of gene transfer into skeletal muscle in vivo, we describe a direct in vitro comparison of two commercially available cationic lipid formulations, Lipofectamine and DOSPER. Optimisation of transfection was performed in the C2C12 mouse muscle cell line, before further studies in primary mouse myoblasts and C2C12 myotubes. Reporter gene constructs expressing either E. coli beta-galactosidase or green fluorescent protein (GFP) were used in order to evaluate transfection efficiency by histochemical staining or FACS analysis, respectively. Both lipid formulations were able to promote efficient, reproducible gene transfer in C2C12 cells, and to transfect primary mouse myoblast cultures successfully. However, DOSPER exhibited the important advantage of being able to transfect cells in the presence of serum of both bovine and murine origin. This feature allowed increased cell survival during in vitro transfections, and may be advantageous for direct in vivo gene transfer efficacy.