Artificial cavitation nuclei significantly enhance acoustically induced cell transfection.
The efficiency of ultrasound-mediated gene transfection was enhanced three- to fourfold, compared to previous results, through the use of green fluorescent protein reporter gene, cultured immortalized human chondrocytes and artificial cavitation nuclei in the form of Albunex. Cells were exposed to 1.0-MHz ultrasound transmitted through the bottom of six-well culture plates containing immortalized chondrocytes, media, DNA at a concentration of 40 micrograms/mL and Albunex at 50 x 10(6) bubbles/mL. Transfection efficiency increased linearly with ultrasound exposure pressure with a transfection threshold observed at a spatial average peak positive pressure (SAPP) of 0.12 MPa and reaching about 50% of the living cells when exposed to 0.41 MPa SAPP for 20 s. Adding fresh Albunex at 50 x 10(6) bubbles/mL prior to sequential 1-s, 0.32- or 0.41-MPa exposures increased transfection with each exposure, reaching 43% transfection after four exposures. Efficient in vitro and in vivo transfection now appear possible with these enhancements.
Greenleaf
W J
WJ
Mayo High School, Rochester, MN, USA.
Bolander
M E
ME
Sarkar
G
G
Goldring
M B
MB
Greenleaf
J F
JF
D000418:Albumins; D002460:Cell Line; D002478:Cells, Cultured; D019902:Chondrocytes; D003287:Contrast Media; D004269:DNA, Bacterial; D005434:Flow Cytometry; D017930:Genes, Reporter / Q000235:genetics*; D006801:Humans; D008863:Microspheres; D010957:Plasmids / Q000235:genetics; D014162:Transfection; D014463:Ultrasonography / Q000379:methods*
D000418:Albumins; C060687:Albunex; D003287:Contrast Media; D004269:DNA, Bacterial