Ultrasound-mediated gene transfection <b>in vitro</b>: Effect of ultrasonic parameters on efficiency and cell viability

Zhang, Yiwei; Tachibana, Rie; Okamoto, Akio; Azuma, Takashi; Sasaki, Akira; Yoshinaka, Kiyoshi; Tei, Yuichi; Takagi, Shu; Matsumoto, Yoichiro

doi:10.3109/02656736.2012.665568

Cited by 13 publications

(12 citation statements)

References 44 publications

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“…There is a considerable heterogeneity in the literature concerning sonoporation devices and procedures, leading to marked differences in transfection efficiency [ 10 , 23 – 25 ]. Another limitation is the fact that most devices are not user-friendly and therefore not adapted for routine use in research laboratories [ 17 , 25 – 26 ]. Some investigators have obtained improved transfection efficiency and chemotherapeutic drug delivery using ultrasound by combining this approach with contrast agents [ 23 – 29 ].…”

Section: Introductionmentioning

confidence: 99%

Spatial and Temporal Control of Cavitation Allows High In Vitro Transfection Efficiency in the Absence of Transfection Reagents or Contrast Agents

et al. 2015

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Sonoporation using low-frequency high-pressure ultrasound (US) is a non-viral approach for in vitro and in vivo gene delivery. In this study, we developed a new sonoporation device designed for spatial and temporal control of ultrasound cavitation. The regulation system incorporated in the device allowed a real-time control of the cavitation level during sonoporation. This device was evaluated for the in vitro transfection efficiency of a plasmid coding for Green Fluorescent Protein (pEGFP-C1) in adherent and non-adherent cell lines. The transfection efficiency of the device was compared to those observed with lipofection and nucleofection methods. In both adherent and non-adherent cell lines, the sonoporation device allowed high rate of transfection of pEGFP-C1 (40–80%), as determined by flow cytometry analysis of GFP expression, along with a low rate of mortality assessed by propidium iodide staining. The transfection efficiency and toxicity of sonoporation on the non-adherent cell lines Jurkat and K562 were similar to those of nucleofection, while these two cell lines were resistant to transfection by lipofection. Moreover, sonoporation was used to produce three stably transfected human lymphoma and leukemia lines. Significant transfection efficiency was also observed in two fresh samples of human acute myeloid leukemia cells. In conclusion, we developed a user-friendly and cost-effective ultrasound device, well adapted for routine in vitro high-yield transfection experiments and which does not require the use of any transfection reagent or gas micro-bubbles.

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Section: Introductionmentioning

confidence: 99%

Spatial and Temporal Control of Cavitation Allows High In Vitro Transfection Efficiency in the Absence of Transfection Reagents or Contrast Agents

et al. 2015

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“…Three mechanical actions can be distinguished: (i) Radiation force action; (ii) pressure interaction with the cell membrane mechanosensors; and (iii) cavitation, which is stable if the bubble just oscillates and inertial if it collapses. Cavitation effects in in vitro cultures have been reported previously [ 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 ]. Changes produced in the cells include membrane disruption, opening of reversible or irreversible pores, ion channels, and toxicity.…”

Section: Introductionmentioning

confidence: 80%

Effects of Non-thermal Ultrasound on a Fibroblast Monolayer Culture: Influence of Pulse Number and Pulse Repetition Frequency

Peñacoba

Gutiérrez

Barrio

et al. 2021

Sensors

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Despite the use of therapeutic ultrasound in the treatment of soft tissue pathologies, there remains some controversy regarding its efficacy. In order to develop new treatment protocols, it is a common practice to carry out in vitro studies in cell cultures before conducting animal tests. The lack of reproducibility of the experimental results observed in the literature concerning in vitro experiments motivated us to establish a methodology for characterizing the acoustic field in culture plate wells. In this work, such acoustic fields are fully characterized in a real experimental configuration, with the transducer being placed in contact with the surface of a standard 12-well culture plate. To study the non-thermal effects of ultrasound on fibroblasts, two different treatment protocols are proposed: long pulse (200 cycles) signals, which give rise to a standing wave in the well with the presence of cavitation (ISPTP max = 19.25 W/cm2), and a short pulse (five cycles) of high acoustic pressure, which produces a number of echoes in the cavity (ISPTP = 33.1 W/cm2, with Pmax = 1.01 MPa). The influence of the acoustic intensity, the number of pulses, and the pulse repetition frequency was studied. We further analyzed the correlation of these acoustic parameters with cell viability, population, occupied surface, and cell morphology. Lytic effects when cavitation was present, as well as mechanotransduction reactions, were observed.

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“…Among the physical effects of ultrasound (US), sonoporation is considered the most important and intriguing. Briefly, as the US waves propagate through the medium, the rapid growth in size (rarefaction) and the collapse of microbubbles (compression) lead to the implosion of the microbubbles (Sirsi and Borden, 2012;Zhang et al, 2012). Formation of shock waves, bubble wall motion, and microjets provide the energy for sonoporation of cell membrane.…”

Section: Discussionmentioning

confidence: 99%

Ultrasound Microbubble-Mediated Delivery of Integrin-Linked Kinase Gene Improves Endothelial Progenitor Cells Dysfunction in Pre-Eclampsia

Cui

Yan²,

Luo³

et al. 2014

DNA and Cell Biology

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Pre-eclampsia (PE) is a specific vascular complication in pregnancy whose precise mechanism is still unclear. We hypothesized that endothelial progenitor cells (EPCs), the precursor of endothelial cells, might be impaired in patients with PE and hold a great promise for the treatment of PE. In the present study, we analyzed the EPCs number and expression of integrin-linked kinase (ILK) in PE patients. We confirmed that both EPCs number and ILK expression were diminished in PE patients. Next, we transfected EPCs with ILK gene using ultrasonic microbubble technique (UMT) for the first time, as UMT is a novel type of gene transfer technology showing promising applications in stem cells apart from EPCs. To further investigate the transfection efficiency of UMT, RT-PCR analysis and western blot were used to examine the messenger RNA (mRNA) and protein level of ILK. After transfection of the ILK gene, EPCs function was tested to illustrate the role of ILK in cell proliferation, apoptosis, migration, and secretion. The results of the in vitro study suggested that UMT, a novel gene delivery system, could be considered a potent physical method for EPCs transfection. Moreover, the growth and angiogenetic properties of EPCs are enhanced by introducing ILK. This study may afford a new trend for EPCs transfection and gene therapy in PE.

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Ultrasound-mediated gene transfection in vitro: Effect of ultrasonic parameters on efficiency and cell viability

Cited by 13 publications

References 44 publications

Spatial and Temporal Control of Cavitation Allows High In Vitro Transfection Efficiency in the Absence of Transfection Reagents or Contrast Agents

Spatial and Temporal Control of Cavitation Allows High In Vitro Transfection Efficiency in the Absence of Transfection Reagents or Contrast Agents

Effects of Non-thermal Ultrasound on a Fibroblast Monolayer Culture: Influence of Pulse Number and Pulse Repetition Frequency

Ultrasound Microbubble-Mediated Delivery of Integrin-Linked Kinase Gene Improves Endothelial Progenitor Cells Dysfunction in Pre-Eclampsia

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