Analysis of in vitro Transfection by Sonoporation Using Cationic and Neutral Microbubbles

Tlaxca, Jose L.; Anderson, Christopher R.; Klibanov, Alexander L.; Lowrey, Bryce; Hossack, John A.; Alexander, J. Steven; Lawrence, Michael B.; Rychak, Joshua J.

doi:10.1016/j.ultrasmedbio.2010.05.014

« Previous Next »Ultrasound in Medicine and Biology
Volume 36, Issue 11 , Pages 1907-1918, November 2010

Analysis of in vitro Transfection by Sonoporation Using Cationic and Neutral Microbubbles

Jose L. Tlaxca
- Department of Biomedical Engineering, University of Virginia, Charlottesville, VA
Christopher R. Anderson
- Targeson, Inc., San Diego, CA
Alexander L. Klibanov
- Department of Biomedical Engineering, University of Virginia, Charlottesville, VA
- Cardiovascular Research Center, University of Virginia, Charlottesville, VA
- Cardiovascular Division, Department of Internal medicine, University of Virginia, Charlottesville, VA
Bryce Lowrey
- Department of Biomedical Engineering, University of Virginia, Charlottesville, VA
John A. Hossack
- Department of Biomedical Engineering, University of Virginia, Charlottesville, VA
J. Steven Alexander
- Department of Molecular and Cellular Physiology, Louisiana State University Health Science center, Shreveport, LA
Michael B. Lawrence
- Department of Biomedical Engineering, University of Virginia, Charlottesville, VA
- Address correspondence to: Michael B. Lawrence, Ph.D., Associate Professor and Chair, Department of Biomedical Engineering, University of Virginia, MR-5 Building, Room 2111, P.O. Box 800759, Charlottesville, VA 22908.
Joshua J. Rychak
- Targeson, Inc., San Diego, CA

Received 17 March 2010; received in revised form 13 May 2010; accepted 14 May 2010. published online 01 September 2010.

Abstract

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/cm² 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. (E-mail: mbl2a@virginia.edu)

Key Words: Ultrasound, Sonoporation, Microbubble, Transfection efficiency, Gene delivery, Cationic microbubbles