Analysis of in vitro transfection by sonoporation using cationic and neutral microbubbles.
The objective of the study was to examine the role of acoustic power intensity and microbubble and plasmid concentrations on transfection efficiency in HEK-293 cells using a sonoporator with a 1-MHz transducer. A green fluorescent protein (GFP) reporter plasmid was delivered in as much as 80% of treated cells, and expression of the GFP protein was observed in as much as 75% of cells, using a power intensity of 2 W/cm(2) with a 25% duty cycle. In addition, the relative transfection abilities of a lipid noncationic and cationic microbubble platform were investigated. As a positive control, cells were transfected using Lipofectamine reagent. Cell survival and transfection efficiency were inversely proportional to acoustic power and microbubble concentration. Our results further demonstrated that high-efficiency transfection could be achieved, but at the expense of cell loss. Moreover, direct conjugation of plasmid to the microbubble did not appear to significantly enhance transfection efficiency under the examined conditions, although this strategy may be important for targeted transfection in vivo.
Tlaxca
Jose L
JL
Department of Biomedical Engineering, University of Virginia, Charlottesville, VA 22908, USA.
Anderson
Christopher R
CR
Klibanov
Alexander L
AL
Lowrey
Bryce
B
Hossack
John A
JA
Alexander
J Steven
JS
Lawrence
Michael B
MB
Rychak
Joshua J
JJ
D002412:Cations; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics*; D066298:In Vitro Techniques; D045423:Microbubbles; D008856:Microscopy, Fluorescence; D010957:Plasmids; D014159:Transducers; D014162:Transfection; D014465:Ultrasonics
D002412:Cations; D049452:Green Fluorescent Proteins
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