Comparison of Transfection Efficiency of Nonviral Gene Transfer Reagents

Abstract

This study compared six commercially available reagents (Arrest-In, ExpressFect, FuGENE HD, jetPEI, Lipofectamine 2000, and SuperFect) for gene transfection. We examined the efficiency and cytotoxicity using nine different cell lines (MC3T3-E1 mouse preosteoblasts, PT-30 human epithelial precancer cells, C3H10T1/2 mouse stem cells, MCF-7 human breast cancer cells, HeLa human cervical cancer, C2C12 mouse myoblasts, Hep G2 human hepatocellular carcinoma, 4T1 mouse mammary carcinoma, and HCT116 human colorectal carcinoma), and primary cells (HEKn human epidermal keratinocytes) with two different plasmid DNAs encoding luciferase or β-galactosidase in the presence or absence of serum. Maximal transfection efficiency in MC3T3-E1, C3H10T1/2, HeLa, C2C12, Hep G2, and HCT116 was seen using FuGENE HD, in PT-30, 4T1, and HEKn was seen using Arrest-In, and in MCF-7 was seen using jetPEI. Determination of cytotoxicity showed that the largest amount of viable cells was found after transfection with jetPEI and ExpressFect. These results suggest that FuGENE HD is the most preferred transfection reagent for many cell lines, followed by Arrest-In and jetPEI. These results may be useful for improving nonviral gene and cell therapy applications.

A comparative study on nonviral genetic modifications in cord blood and bone marrow mesenchymal stem cells

“…Such difference might be the result of difference in GFP expression in hUSSCs and hMSCs or size-dependency of nucleic acid transfection in both stem cell types. Altogether it revealed that the nature of the cell itself plays a critical role in effectiveness of transfection as suggested by others (Yamano et al 2010;Lee and Kim 2005).…”

“…These reagents were reported to be efficient in some previous studies (Gheisari et al 2008;Yamano et al 2010;Akimov et al 2009;Nabzdyk et al 2011).…”

“…Few approaches to optimize the efficiency of gene transfer and establishment of proficient transfection protocols have been described only for some types of cells (Dinser et al 2001;Yamano et al 2010;Gonzalez et al 2007;Yamauchi et al 2010). Because of variation in transfection yield and cytotoxicity of each method, selection of an appropriate method for transfection of a given cell type is crucial.…”

“…Many different nonviral methods have been developed to deliver nucleic acid to mammalian cells for both stable and transient expression, including DEAE dextran, commercial non-viral vectors (lipids, polymers, dendrimer, polyethylenimine, peptides, and nanoparticle-based compounds) and electroporation (Yamano et al 2010). However, electroporation suffers from many limitations, including the need for large cell numbers, large amounts of nucleic acid, serum-free transfection conditions, and the high level of cell death (Andre and Mir 2004).…”

“…In general, transfection efficiency of vectors depends on cell types, kind of nucleic acid, and medium conditions (Yamano et al 2010). Although many new systems hold promises to achieve hightransfection efficiencies, a systematic evaluation considering some types of stem cells has not, to our knowledge, been performed yet.…”