Comparative transfection of DNA into primary and transformed mammalian cells from different lineages

<p>Abstract</p> <p>Background</p> <p>The delivery of DNA into human cells has been the basis of advances in the understanding of gene function and the development of genetic therapies. Numerous chemical and physical approaches have been used to deliver the DNA, but thei...

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Main Authors: Bedayat Babak, Emamekhoo Hamid, De Semir David, Maurisse Rosalie, Abdolmohammadi Alireza, Parsi Hooman, Gruenert Dieter C
Format: Article
Language:English
Published: BMC 2010-02-01
Series:BMC Biotechnology
Online Access:http://www.biomedcentral.com/1472-6750/10/9
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author Bedayat Babak
Emamekhoo Hamid
De Semir David
Maurisse Rosalie
Abdolmohammadi Alireza
Parsi Hooman
Gruenert Dieter C
author_facet Bedayat Babak
Emamekhoo Hamid
De Semir David
Maurisse Rosalie
Abdolmohammadi Alireza
Parsi Hooman
Gruenert Dieter C
author_sort Bedayat Babak
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>The delivery of DNA into human cells has been the basis of advances in the understanding of gene function and the development of genetic therapies. Numerous chemical and physical approaches have been used to deliver the DNA, but their efficacy has been variable and is highly dependent on the cell type to be transfected.</p> <p>Results</p> <p>Studies were undertaken to evaluate and compare the transfection efficacy of several chemical reagents to that of the electroporation/nucleofection system using both adherent cells (primary and transformed airway epithelial cells and primary fibroblasts as well as embryonic stem cells) and cells in suspension (primary hematopoietic stem/progenitor cells and lymphoblasts). With the exception of HEK 293 cell transfection, nucleofection proved to be less toxic and more efficient at effectively delivering DNA into the cells as determined by cell proliferation and GFP expression, respectively. Lipofectamine and nucleofection of HEK 293 were essentially equivalent in terms of toxicity and efficiency. Transient transfection efficiency in all the cell systems ranged from 40%-90%, with minimal toxicity and no apparent species specificity. Differences in efficiency and toxicity were cell type/system specific.</p> <p>Conclusions</p> <p>In general, the Amaxa electroporation/nucleofection system appears superior to other chemical systems. However, there are cell-type and species specific differences that need to be evaluated empirically to optimize the conditions for transfection efficiency and cell survival.</p>
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spelling doaj.art-3c3dd34f782f4a1485d582f73c1008982022-12-21T19:14:09ZengBMCBMC Biotechnology1472-67502010-02-01101910.1186/1472-6750-10-9Comparative transfection of DNA into primary and transformed mammalian cells from different lineagesBedayat BabakEmamekhoo HamidDe Semir DavidMaurisse RosalieAbdolmohammadi AlirezaParsi HoomanGruenert Dieter C<p>Abstract</p> <p>Background</p> <p>The delivery of DNA into human cells has been the basis of advances in the understanding of gene function and the development of genetic therapies. Numerous chemical and physical approaches have been used to deliver the DNA, but their efficacy has been variable and is highly dependent on the cell type to be transfected.</p> <p>Results</p> <p>Studies were undertaken to evaluate and compare the transfection efficacy of several chemical reagents to that of the electroporation/nucleofection system using both adherent cells (primary and transformed airway epithelial cells and primary fibroblasts as well as embryonic stem cells) and cells in suspension (primary hematopoietic stem/progenitor cells and lymphoblasts). With the exception of HEK 293 cell transfection, nucleofection proved to be less toxic and more efficient at effectively delivering DNA into the cells as determined by cell proliferation and GFP expression, respectively. Lipofectamine and nucleofection of HEK 293 were essentially equivalent in terms of toxicity and efficiency. Transient transfection efficiency in all the cell systems ranged from 40%-90%, with minimal toxicity and no apparent species specificity. Differences in efficiency and toxicity were cell type/system specific.</p> <p>Conclusions</p> <p>In general, the Amaxa electroporation/nucleofection system appears superior to other chemical systems. However, there are cell-type and species specific differences that need to be evaluated empirically to optimize the conditions for transfection efficiency and cell survival.</p>http://www.biomedcentral.com/1472-6750/10/9
spellingShingle Bedayat Babak
Emamekhoo Hamid
De Semir David
Maurisse Rosalie
Abdolmohammadi Alireza
Parsi Hooman
Gruenert Dieter C
Comparative transfection of DNA into primary and transformed mammalian cells from different lineages
BMC Biotechnology
title Comparative transfection of DNA into primary and transformed mammalian cells from different lineages
title_full Comparative transfection of DNA into primary and transformed mammalian cells from different lineages
title_fullStr Comparative transfection of DNA into primary and transformed mammalian cells from different lineages
title_full_unstemmed Comparative transfection of DNA into primary and transformed mammalian cells from different lineages
title_short Comparative transfection of DNA into primary and transformed mammalian cells from different lineages
title_sort comparative transfection of dna into primary and transformed mammalian cells from different lineages
url http://www.biomedcentral.com/1472-6750/10/9
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AT maurisserosalie comparativetransfectionofdnaintoprimaryandtransformedmammaliancellsfromdifferentlineages
AT abdolmohammadialireza comparativetransfectionofdnaintoprimaryandtransformedmammaliancellsfromdifferentlineages
AT parsihooman comparativetransfectionofdnaintoprimaryandtransformedmammaliancellsfromdifferentlineages
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