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J Biomol Tech. 2017 Jul;28(2):67-74. doi: 10.7171/jbt.17-2802-003. Epub 2017 Apr 29.

Comparative Analysis of Non-viral Transfection Methods in Mouse Embryonic Fibroblast Cells.

Author information

1
Department of Medicine, Boston University School of Medicine, Boston, Massachusetts 02118, USA.

Abstract

Mouse embryonic fibroblast (MEF) cells are an important in vitro model for developmental biology, disease, and reprogramming studies. However, as with other primary cells, they are challenging to transfect. Although viral gene-delivery methods achieve high gene-delivery efficiency, challenges with cell mutagenesis and safety among others have led to the use and improvement of non-viral gene-delivery methods in MEF cells. Despite the importance of gene delivery in MEF cells, there is limited comparison of method/reagent efficacy. In this study, we compared the effectiveness of different gene-delivery methods and several reagents currently available in MEF cells by introducing a plasmid containing enhanced green fluorescent protein (EGFP). We analyze transfection efficiency by EGFP fluorescence. Our results suggest that two gene-delivery methods-electroporation and magnetofection in combination with a lipid reagent, are the most efficient transfection methods in MEF cells. This study provides a foundation for the selection of transfection methods or reagents when using MEF cells.

KEYWORDS:

MEF; dendrimer; electroporation; lipid-based transfection; magnetofection

PMID:
28507462
PMCID:
PMC5410126
DOI:
10.7171/jbt.17-2802-003
[Indexed for MEDLINE]
Free PMC Article

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