Lipid-mediated gene transfer of acidic fibroblast growth factor into human corneal endothelial cells
Lipid-mediated gene transfer of acidic fibroblast growth factor into human corneal endothelial cells
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DOI：
10.1016/j.exer.2004.08.024
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发表时间：
2005-01-01
影响因子：
3.4
通讯作者：
Pleyer, U
中科院分区：
文献类型：
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作者：
Dannowski, H;Bednarz, J;Pleyer, U
本研究旨在优化非病毒基因转染条件，并探讨成纤维细胞生长因子-1（FGF-1）基因转染对人角膜内皮细胞（HCEC）增殖的影响。使用5种非病毒载体（Lipofectin（TM）、DMRIE-C（TM）、DAC-30、Atriotene（TM）、FuGene（TM）6）与编码增强型绿色荧光蛋白（EGFP）和FGF-1的质粒转染HCEC。通过FRC分析使用EGFP构建体定量和优化转染效率和毒性（n = 6）。使用最佳条件下，HCEC转染FGF-1质粒和细胞增殖以及FGF-1的表达，分别在第4和第7天通过计数和蛋白质印迹法测定。Lipofectin（17 +/- 2.02%）转染HCEC的成功率高于DMRIE-C（11 +/- 1.46%）、异戊烯（9 +/- 0.62%）、FuGene（9 +/- 0.93%）和DAC-30（7 +/- 0.59%）。脂质的毒性范围为2%至4%。DAC-30/FGF-1对HCEC的增殖作用最佳（P
The aim of this study was to optimize non-viral gene transfer conditions and investigate the effect of fibroblast growth factor-1 (FGF-1) gene transfer on human corneal endothelial cell (HCEC) proliferation. Five non-viral vectors (Lipofectin(TM), DMRIE-C(TM), DAC-30, Effectene(TM), FuGene(TM)6) were used to transfect HCEC with plasmids coding for enhanced green fluorescent protein (EGFP) and FGF-1. Transfection efficiency and toxicity (n = 6) were quantified and optimized using the EGFP construct by FRCS-analysis. Using optimal conditions HCEC were transfected with the FGF-1 plasmid and cell proliferation as well as expression of FGF-1 were determined at days 4 and 7 by counting and western blotting, respectively. Lipofectin (17 +/- 2.02%) transfected HCEC more successfully than DMRIE-C (11 +/- 1.46%), Effectene (9 +/- 0.62%), FuGene (9 +/- 0.93%) and DAC-30 (7 +/- 0.59%). Toxicity of the lipids ranged from 2 to 4%. Optimal HCEC proliferation was achieved with DAC-30/FGF-1 (P