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Comparison of transfection efficiency of nonviral gene transfer reagents..pdf

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Comparison of transfection efficiency of nonviral gene transfer reagents.

RESEARCH Comparison of Transfection Efficiency of Nonviral Gene Transfer Reagents Seiichi Yamano ? Jisen Dai ? Amr M. Moursi Published online: 29 June 2010 Springer Science+Business Media, LLC 2010 Abstract This study compared six commercially avail- able 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 preos- teoblasts, PT-30 human epithelial precancer cells, C3H10T1/2 mouse stem cells, MCF-7 human breast cancer cells, HeLa human cervical cancer, C2C12 mouse myo- blasts, Hep G2 human hepatocellular carcinoma, 4T1 mouse mammary carcinoma, and HCT116 human colo- rectal carcinoma), and primary cells (HEKn human epi- dermal keratinocytes) with two different plasmid DNAs encoding luciferase or b-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. Keywords Gene transfection Cationic lipid Polymer Polyethylenimine Activated-dendrimer Introduction Gene transfection is a widely used technique for molecular studies and therapeutics. Many of vectors have been developed with the purpose of delivering genes efficiently into target cells and protecting them from nuclease deg- radation. Viral vectors are generally most efficient for cell transduction, but they present several disadvantages related with immunogenicity, inflammation, DNA size restriction, and large-scale cons