Induction of apoptosis in sonoporation and ultrasonic gene transfer.

Ultrasound in Medicine & Biology
Douglas L Miller, Chunyan Dou

Abstract

The role of apoptosis in sonoporation and ultrasound-enhanced gene transfection of cell suspensions was examined in vitro. Suspensions of HL-60 and of CHO-K1 cells were exposed to 2.25-MHz continuous ultrasound for 1 min in a 60-rpm rotating-tube exposure system, with ultrasound contrast media added to ensure nucleation of cavitation. Cell necrosis was measured by trypan blue dye exclusion (using a hemacytometer) and by propidium iodide nuclear staining (using flow cytometry). Apoptosis was detected by the annexin V method with Alexa Fluor 350 as the fluorescent label, and confirmed by Hoechst 33342 nuclear staining. Sonoporation cell loading was assessed by uptake of large fluorescent-dextran molecules from the medium. Transfection was demonstrated by expression of green fluorescent protein (GFP) from plasmids transferred into the cells by the treatment. Cell scoring was performed by flow cytometry, with necrotic cell events excluded. For HL-60 cells at 0.4 MPa, cell loading and transfection was significantly increased relative to shams at 2, 6 and 24 h post exposure, peaking at 19.0 +/- 5.5% and 9.6 +/- 4.2% of non-necrotic cells, respectively, at 6 h. However, about one third of the treatment-positive cells were identified a...Continue Reading

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Citations

Nov 18, 2011·The Journal of the Acoustical Society of America·Douglas L Miller, Chunyan Dou
Aug 3, 2013·Advanced Drug Delivery Reviews·Helen MulvanaMartyn Hill
Jun 5, 2013·Molecular Therapy : the Journal of the American Society of Gene Therapy·Misty L NobleCarol H Miao
Apr 1, 2011·Journal of Medical Ultrasonics·Mariame A HassanTakashi Kondo
Mar 12, 2016·Ultrasonics Sonochemistry·Paras JawaidTakashi Kondo
Aug 27, 2013·Ultrasound in Medicine & Biology·Eun Sun LeeByung Ihn Choi
Aug 28, 2012·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Mariame A HassanTakashi Kondo
Aug 28, 2012·American Journal of Surgery·Ahmed AlzaraaDavid M Lloyd
Apr 17, 2012·Ultrasound in Medicine & Biology·Peng QinAlfred C H Yu
Oct 29, 2011·Ultrasound in Medicine & Biology·Wenjing ZhongAlfred C H Yu
Oct 4, 2011·Ultrasound in Medicine & Biology·Laura KowalczukPascal Laugier
Aug 31, 2010·Ultrasound in Medicine & Biology·Jose L TlaxcaJoshua J Rychak
Jul 27, 2010·Ultrasonics·Kelsey J Carvell, Timothy A Bigelow
Mar 31, 2010·Ultrasound in Medicine & Biology·Robyn K SchlicherMark R Prausnitz
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Sep 16, 2017·Journal of Biomedical Optics·Paulius RuzgysSaulius Šatkauskas
Mar 6, 2021·Scientific Reports·Xinxing DuanAlfred C H Yu

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