Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2012;46(6):347-353.
doi: 10.3103/S0095452712060035. Epub 2012 Dec 5.

Optimization of transfection methods for Huh-7 and Vero cells: A comparative study

A Hashemi  1 , F Roohvand  1   2 , M H Ghahremani  3 , M R Aghasadeghi  1 , R Vahabpour  1 , F Motevali  1   2 , A Memarnejadian  1
Affiliations

Optimization of transfection methods for Huh-7 and Vero cells: A comparative study

A Hashemi et al. Cytol Genet. 2012.

Abstract

Availability of an efficient transfection protocol is the first determinant in success of gene transferring studies in mammalian cells which is accomplished experimentally for every single cell type. Herein, we provide data of a comparative study on optimization of transfection condition by electroporation and chemical methods for Huh-7 and Vero cells. Different cell confluencies, DNA/reagent ratios and total transfection volumes were optimized for two chemical reagents including jetPEI™ and Lipofectamine™ 2000. Besides, the effects of electric field strength and pulse length were investigated to improve electroporation efficiency. Transfection of cells by pEGFP-N1 vector and tracking the expression of GFP by FACS and Fluorescence Microscopy analysis were the employed methods to evaluate transfection efficiencies. Optimized electroporation protocols yielded 63.73 ± 2.36 and 73.9 ± 1.6% of transfection in Huh-7 and Vero cells respectively, while maximum achieved level of transfection by jetPEI™ was 14.2 ± 0.69 and 28 ± 1.11% Huh-7 and Vero cells, respectively. Post transfectional chilling of the cells did not improve electrotransfection efficiency of Huh-7 cells. Compared to chemical based reagents, electroporation showed superior levels of transfection in both cell lines. The presented protocols should satisfy most of the experimental applications requiring high transfection efficiencies of these two cell lines.

Keywords: Cationic Lipid; Electric Field Strength; Infectious Bronchitis Virus; Transfection Efficiency; Vero Cell.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

References

    1. Kim T.K., Eberwine J.H. Mammalian Cell Transfection: the Present and the Future. Anal. Bioanal. Chem. 2010;397:3173–3178. doi: 10.1007/s00216-010-3821-6. - DOI - PMC - PubMed
    1. Recillas-Targa F. Multiple Strategies for Gene Transfer, Expression, Knockdown, and Chromatin Influence in Mammalian Cell Lines and Transgenic Animals. Mol. Biotechnol. 2006;34:337–354. doi: 10.1385/MB:34:3:337. - DOI - PubMed
    1. Shabani M., Hemmati Sh., Hadavi R., et al. Optimization of Gene Transfection in Murine Myeloma Cell Lines using Different Transfection Reagents, Avicenna. J. Med. Biotech. 2010;2:123–130. - PMC - PubMed
    1. Maurisse R., Semir D.D., Emamekhoo H., et al. Comparative Transfection of DNA into Primary and Transformed Mammalian Cells from Different Lineages. BMC Biotechnol. 2010;10:2–9. doi: 10.1186/1472-6750-10-9. - DOI - PMC - PubMed
    1. Colosimo A., Goncz K.K., Holmes A.R., et al. Transfer and Expression of Foreign Genes in Mammalian Cells. BioTechniques. 2000;29:314–331. - PubMed

LinkOut - more resources