Astrocytic responses to DNA delivery using nucleofection.
Nucleofection is a powerful non-viral transfection technique that can deliver plasmid DNA with high efficiency to cells that are traditionally difficult to transfect. In this study, we demonstrate that nucleofection of astrocytes grown in primary cell culture resulted in 76 ± 9% transfected cells and low cytotoxicity. However, the nucleofected astrocytes showed a reduced re-attachment to the growth media when replated and subsequent impairment of proliferation. This led to substantially decreased cell densities during the initial 72 h following transfection. Furthermore, these cells were less efficient at producing wound closure in a scratch model of injury. Nucleofection also resulted in the generation of a small proportion of polynucleated cells. The findings demonstrate that nucleofection provides a valuable technique for delivering DNA to astrocytes in culture. However, considerable care is needed in designing and interpreting such studies because of long-lasting changes induced in key properties of these cells by the nucleofection process.
Muyderman
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H
Centre for Neuroscience, Flinders Medical Science and Technology, School of Medicine, Flinders University, Adelaide, SA, Australia. hakan.muyderman@flinders.edu.au
Yew
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Homkajorn
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B
Sims
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D000818:Animals; D001253:Astrocytes / Q000166:cytology*; D049109:Cell Proliferation; D004247:DNA / Q000008:administration & dosage*; D018274:Electroporation / Q000379:methods; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000379:methods*; D014945:Wound Healing / Q000502:physiology
D005904:Glial Fibrillary Acidic Protein; D004247:DNA