Nonviral transfection of suspension cells in ultrasound standing wave fields.
Ultrasound-induced cavitation has been widely used for delivering DNA vectors into cells. However, this approach may seriously disrupt cell membranes and cause lethal damage when cells are exposed to the inertial cavitation field. In this study, instead of using sonoporation, ultrasound standing wave fields (USWF) were explored for nonviral transfection of suspension cells. Acoustic resonance in a tubular chamber was generated from the interference of waves emitted from a piezoelectric transducer and consequently reflected from a borosilicate glass coverslip. The suspended K562 erythroleukemia cells were transfected by polyethyleneimine (PEI)/DNA complexes with and without exposure to 1-MHz USWF for 5 min. During USWF exposure, K562 cells moved to the pressure nodal planes first and formed cell bands by the primary radiation force. Nanometer-sized PEI/DNA complexes, circulated between nodal planes by acoustic microstreaming, then used the cell agglomerates as the nucleating sites on which to attach. After incubation at 37 degrees C for 48 h, the efficiency of nonviral transfection based on EGFP transgene expression was determined by fluorescent microscopy and fluorometry. Both studies showed that USWF brought suspended K562 cells and PEI/DNA complexes into close contact at the pressure nodal planes, yielding an approximately 10-fold increment of EGFP transgene expression compared with the group without ultrasonic treatment.
Lee
Yu-Hsiang
YH
Department of Chemical Engineering and Materials Science, University of Southern California, Los Angeles, USA.
Peng
Ching-An
CA
D002470:Cell Survival / Q000235:genetics; D004247:DNA / Q000235:genetics; D005456:Fluorescent Dyes; D005470:Fluorometry / Q000379:methods; D015870:Gene Expression / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D020014:K562 Cells; D008856:Microscopy, Fluorescence / Q000379:methods; D010957:Plasmids / Q000235:genetics; D011094:Polyethyleneimine; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics; D014465:Ultrasonics
D005456:Fluorescent Dyes; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D004247:DNA