Optimization of N2a cell transfection mediated by liposome

doi:10.3969/j.issn.2095-4344.2014.29.014

Abstract

Abstract:

BACKGROUND: Cationic liposome-mediated cell transfection is reliable and repeatable. However the transfection efficiency is often low.

OBJECTIVE: To study the optimized methods for gene transfection mediated by liposome into N2a cells (mouse neuroblastma cells).

METHODS: Using traditional adherent method and improved suspension method, 500 ng recombinant plasmid pcDNA3-GFP carrying green fluorescence protein was transfected into N2a cells in 24-well culture plate, which was mediated by 1.5 μL Lipofectamine™ LTX Reagent. The expression of green fluorescent protein was observed by inverted fluorescence microscope, and the transfection efficiencies at different transfection ways were calculated. By using improved suspension transfection method, 500 ng plasmid DNA was transfected with different doses of Lipofectamine™ LTX Reagent (1.0, 1.5, 2.0, 2.5 μL). The optimal ratio of liposome and DNA was explored.

RESULTS AND CONCLUSION: The transfection efficiency of suspension transfection method was significantly higher than that of the tranditional adherent method (P < 0.01) when using 1.5 μL liposome/500 ng DNA. The transfection efficiency of the 1.0, 1.5, 2.0, 2.5 μL Lipofectamine™ LTX on 500 ng plasmid DNA was respectively (76.60±3.85)%, (80.00±4.17)%, (88.00±5.89)%, (54.96±4.23)%. It showed the 500 ng DNA and 2.0 μL liposome achieve the highest transfection efficiency.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: liposome, transfection, green fluorescent protein

CLC Number:

R318

Zhao Yun-he, Wang Ruo-nan, Yang Gui-jiao, Lu Li. Optimization of N2a cell transfection mediated by liposome[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(29): 4669-4674.

Figures/Tables 2

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