Analysis of in vitro transfection by sonoporation using cationic and neutral microbubbles.

Analysis of in vitro transfection by sonoporation using cationic and neutral microbubbles.
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DOI:
10.1016/j.ultrasmedbio.2010.05.014
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发表时间:
2010-11
影响因子:
2.9
通讯作者:
Rychak, Joshua J.
Rychak, Joshua J.
中科院分区:
医学3区
文献类型:
--
作者:
Tlaxca, Jose L.;Anderson, Christopher R.;Klibanov, Alexander L.;Lowrey, Bryce;Hossack, John A.;Alexander, J. Steven;Lawrence, Michael B.;Rychak, Joshua J.

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本研究的目的是研究声功率强度和微泡和质粒浓度对HEK-293细胞转染效率的作用,使用1 MHz换能器的sonoporator。绿色荧光蛋白(GFP)报告质粒在多达80%的处理细胞中递送,并且在多达75%的细胞中观察到GFP蛋白的表达,使用2 W/cm 2的功率强度和25%的占空比。此外,脂质非阳离子和阳离子微泡平台的相对转染能力进行了研究。作为阳性对照,使用Lipofectamine试剂转染细胞。细胞存活率和转染效率与声功率和微泡浓度成反比。我们的结果进一步证明,可以实现高效转染,但以细胞损失为代价。此外,质粒与微泡的直接缀合似乎没有显着提高转染效率在检查的条件下,虽然这种策略可能是重要的体内靶向转染。
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.
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