Volume 12, Issue 4 (Jul-Aug 2018)                   mljgoums 2018, 12(4): 12-16 | Back to browse issues page


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Gray Z, Douzandegan Y, Tabarraei A, Moradi A. Optimization of SW480 Colon Cancer Cells Transfection with Lipofectamine 2000. mljgoums. 2018; 12 (4) :12-16
URL: http://goums.ac.ir/mljgoums/article-1-1088-en.html
1- Department of Microbiology, school of medicine, Golestan University of medical science, Gorgan, Iran
2- Golestan research center of Gastroenterology and Hepatology, Department of Microbiology, school of medicine, Golestan University of medical science, Gorgan, Iran , abmoradi@gmail.com
Abstract:   (3693 Views)
ABSTRACT
          Background and Objectives: Nonviral carriers including those based on synthetic cationic lipids, offer several advantages over the viral counterparts. These carriers are able to form complexes with nucleic acids and deliver genes into the cells via the cellular endocytosis pathway, without significant toxicity. The level of transgenes expression depends on some experimental variables including cell type and density, Lipofectamine and DNA concentrations and Lipofectamine-DNA complexing time. The main objective of this study was to optimize transfection of SW480 colon cancer cells with Lipofectamine 2000.
          Methods: In this study, SW480 cells were transfected with plasmid containing green fluorescent protein reporter gene using Lipofectamine 2000. Green fluorescent protein expression was studied under a reverse fluorescence microscope and the results were analyzed with the ImageJ software. Effect of different quantities of plasmid DNA and different Lipofectamine 2000 volumes on cell transfection efficiency was evaluated.
          Results: The optimal volume of Lipofectamine and quantity of plasmid was 2 µl and 1µg, respectively, which showed 59% efficiency for the transfection of SW480 cells at 24 hours post-transfection.
          Conclusion: This study shows that Lipofectamine 2000 is an efficient reagent for the delivery of genes into SW480 cells. According to the results, the quantity of DNA per transfection and reagent concentrations are essential factors for a successful transfection.
          Keywords: Optimization; pEGFP-NI; Lipofectamine; SW480.
Full-Text [PDF 596 kb]   (520 Downloads)    
Type of Study: Original Paper |
Received: 2018/05/30 | Accepted: 2018/05/30 | Published: 2018/05/30 | ePublished: 2018/05/30

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