A novel transfection method for mammalian cells using calcium alginate microbeads
A novel transfection method for mammalian cells using calcium alginate microbeads
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DOI：
10.1016/s1389-1723(04)70189-9
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发表时间：
2004-03-01
影响因子：
2.8
通讯作者：
Fukui, K
中科院分区：
文献类型：
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作者：
Higashi, T;Nagamori, E;Fukui, K
将遗传物质直接转移到哺乳动物细胞中是一项不可或缺的技术。我们研制了海藻酸钙（CA）微球，可将质粒DNA和酵母人工染色体导入植物和酵母细胞。在本文中，我们证明了有效的转染哺乳动物细胞的CA微珠固定质粒DNA。用含有巨细胞病毒（CMV）启动子和增强型绿色荧光蛋白（EGFP）基因的pEGFP-C1质粒进行转染。转染后24 h观察到EGFP的瞬时表达。当海藻酸钠浓度为1%，质粒DNA量增加到100 μ g时，表达效率最高。使用CA微珠的我们的方法的表达效率比聚乙二醇（PEG）方法高2-10倍。我们的研究结果表明，CA微珠介导的哺乳动物细胞的转染有效地将遗传物质传递到哺乳动物悬浮细胞。
The direct transfer of genetic materials into mammalian cells is an indispensable technique. We have developed calcium alginate (CA) microbeads which can deliver plasmid DNAs and yeast artificial chromosomes into plant and yeast cells. In this paper, we demonstrate the effective transfection of mammalian cells by CA microbeads immobilizing plasmid DNAs. The transfection was performed using the pEGFP-C1 plasmid containing the cytomegalovirus (CMV) promoter and enhanced green fluorescent protein (EGFP) gene. The transient expression of EGFP was observed 24 h after transfection. The expression efficiency was maximum when the concentration of sodium alginate was 1% and the amount of plasmid DNA was increased to 100 mug. The expression efficiency of our method using CA microbeads is 2-10 times higher than that of the polyethylene glycol (PEG) method. Our results suggest that the CA microbead mediated transfection of mammalian cells effectively delivers genetic materials into mammalian suspension cells.