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Open Access Highly Accessed Research article

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

Rosalie Maurisse14, David De Semir1, Hamid Emamekhoo12, Babak Bedayat15, Alireza Abdolmohammadi126, Hooman Parsi1 and Dieter C Gruenert123*

Author Affiliations

1 California Pacific Medical Center Research Institute, San Francisco, CA, USA

2 Department of Laboratory Medicine, University of California San Francisco, San Francisco, CA, USA

3 Department of Medicine, University of Vermont, Burlington, VT, USA

4 Current address: Medicen, 6 rue Alexandre Cabanel, 75015 Paris, France

5 Current address: Department of Anesthesiology and Critical Care, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA

6 Current address: Department of Internal Medicine, Good Samaritan Hospital, Cincinnati, OH, USA

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BMC Biotechnology 2010, 10:9  doi:10.1186/1472-6750-10-9

Published: 8 February 2010

Abstract

Background

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.

Results

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.

Conclusions

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.