PEI/DNA formation affects transient gene expression in suspension Chinese hamster ovary cells via a one-step transfection process

Polyethylenimine (PEI) has been used widely in transient gene expression studies of mammalian cells. We performed transient gene expression in suspension Chinese hamster ovary cells using a one-step transfection procedure in which DNA and PEI were simultaneously added to a cell culture in suspension...

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Bibliographic Details
Published in:Cytotechnology (Dordrecht) 2013-03, Vol.65 (2), p.263-271
Main Authors: Xie, Qiuling, Xinyong, Guo, Xianjin, Chen, Yayu, Wang
Format: Article
Language:English
Subjects:
Biochemistry
Biomedicine
Biotechnology
Cell culture
Chemistry
Chemistry and Materials Science
Deoxyribonucleic acid
DNA
Efficiency
Gene expression
Humidity
Lasers
Mammalian cells
Original Research
Ovaries
Plasmids
Polyethyleneimine
Proteins
Transfection
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Summary:Polyethylenimine (PEI) has been used widely in transient gene expression studies of mammalian cells. We performed transient gene expression in suspension Chinese hamster ovary cells using a one-step transfection procedure in which DNA and PEI were simultaneously added to a cell culture in suspension without prior PEI/DNA complex incubation. To further understand the effect of PEI/DNA formation on the transfection and expression of exogenous gene in shaking state, we investigated the diameter and overcharge of the PEI/DNA complex. The results showed that the diameter of the complex was smaller with more positive charge when the PEI/DNA ratio was higher. Moreover, DNA more easily penetrated cells and nuclei at higher PEI concentrations. The highest transcription level, transfection efficiency, and GFP expression were obtained when the PEI/DNA ratio was 5:1.
ISSN:0920-9069
1573-0778
DOI:10.1007/s10616-012-9483-9