TAILIEUCHUNG - Báo cáo sinh học: "Electroporation by nucleofector is the best nonviral transfection technique in human endothelial and smooth muscle cells"

Tuyển tập các báo cáo nghiên cứu về sinh học được đăng trên tạp chí sinh học Journal of Biology đề tài: Electroporation by nucleofector is the best nonviral transfection technique in human endothelial and smooth muscle cells | Genetic Vaccines and Therapy BioMed Central Methodology Open Access Electroporation by nucleofector is the best nonviral transfection technique in human endothelial and smooth muscle cells Nina Iversen Baard Birkenes Kari Torsdalen and Srdjan Djurovic Address Department of Medical Genetics Ullevâl University Hospital Oslo Norway Email Nina Iversen - Baard Birkenes - Kari Torsdalen - Srdjan Djurovic - Corresponding author Published 18 April 2005 Received 07 December 2004 Genetic Vaccines and Therapy 2005 3 2 doi 1479-0556-3-2 Accepted 18 April 2005 This article is available from http content 3 1 2 2005 Iversen et al licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License http licenses by which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. Abstract Background The aim of this study was to determine the optimal non-viral transfection method for use in human smooth muscle cells SMC and endothelial cells EC . Methods Coronary Artery CoA and Aortic Ao SMC and EC were transfected with a reporter plasmid encoding chloramphenicol acetyltransferase type 1 CAT with seven different transfection reagents two electroporation methods and a photochemical internalization PCI method. CAT determination provided information regarding transfection efficiency and total protein measurement was used to reflect the toxicity of each method. Results Electroporation via the nucleofector machine was the most effective method tested. It exhibited a 10 to 20 fold for SMC and EC respectively increase in transfection efficiency in comparison to the lipofection method combined with acceptable toxicity. FuGene 6 and Lipofectamine PLUS were the preferred transfection reagents tested and .

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