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

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 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 to 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.

Author: Rosalie MaurisseDavid De SemirHamid EmamekhooBabak BedayatAlireza AbdolmohammadiHooman ParsiDieter Gruenert
Credits/Source: BMC Biotechnology 2010, 10:9

Published on: 2010-02-08

Copyright by the authors listed above - made available via BioMedCentral (Open Access). Please make sure to read our disclaimer prior to contacting 7thSpace Interactive. To contact our editors, visit our online helpdesk. If you wish submit your own press release, click here.

Comments Page 0 of 0

There are currently 0 comments to display.

+ Add New Comment

Custom Search