In vitro and in vivo transfection efficiency of a novel ultradeforinable cationic liposome
In vitro and in vivo transfection efficiency of a novel ultradeforinable cationic liposome
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DOI：
10.1016/s0142-9612(03)00534-9
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发表时间：
2004-01-01
期刊：
影响因子：
14
通讯作者：
Kim, CK
中科院分区：
文献类型：
--
作者：
Kim, A;Lee, EH;Kim, CK
阳离子脂质经常被用作制造最有前途的非病毒基因传递系统的主要成分之一，而胆酸钠，一种被称为边缘激活剂的表面活性剂，已被用于制备超可形成和超柔韧性的脂质体，称为转移体。以阳离子脂质体、DOTAP和胆酸钠为原料，制备了一种新型的超成形阳离子脂质体（UCL）。该配方的平均粒径约为80纳米。在不同温度（4℃和20℃）下测定60天的理化稳定性。对新配方的超成形性进行了评价，证明了该配方是可变形的。通过绿色荧光蛋白（GFP）在OVCAR-3（人卵巢癌细胞）、HepG2（人肝癌细胞）、H-1299（人肺癌细胞）和T98G（人脑癌细胞）四种细胞系中的表达，评估质粒DNA/UCL的体外转染效率。除人脑癌细胞外，所有细胞系中DNA与脂质体的最佳转染率为1:14 （w/w）。通过非侵入性地将DNA/UCL复合物应用于小鼠脱毛的背部皮肤，测试了同一配方的体内转染效率及其在器官内的保留时间。研究发现，一旦在完整的皮肤上涂抹，基因可以在6天内转运到几个器官中。(C) 2003 Elsevier Ltd.版权所有。
Cationic lipids have been often used as one of the major components in making most promising non-viral gene delivery systems, whereas sodium cholate, a surfactant so-called edge activator has been used in preparing ultradeformable and ultraflexible liposomes called Transfersomes. Using both a cationic lipid, DOTAP and sodium cholate, a novel formulation of ultradeformable cationic liposome (UCL) has been prepared. The average particle size of this formulation was approximately 80 nm. The physical and chemical stabilities at two different temperatures (4degreesC and 20degreesC) were also evaluated for 60 days. The ultradeformability of new formulation was also assessed, and it has been proved that the formulation is deformable. In vitro transfection efficiency of plasmid DNA/UCL was assessed by the expression of green fluorescent protein (GFP) in four cell lines, OVCAR-3 (human ovarian carcinoma cells), HepG2 (human hepatoma cells), H-1299 (human lung carcinoma cells) and T98G (human brain carcinoma cells). The optimal ratio of DNA to liposome for maximal transfection efficiency was 1:14 (w/w) in all the cell lines except for the human brain carcinoma cells. The same formulation was tested for in vivo transfection efficiency and its retention time within the organs by applying the DNA/UCL complexes on hair-removed dorsal skin of mice non-invasively. It was found that genes were transported into several organs for 6 days once applied on intact skin. (C) 2003 Elsevier Ltd. All rights reserved.