Microfluidic electro-sonoporation: a multi-modal cell poration methodology through simultaneous application of electric field and ultrasonic wave
Microfluidic electro-sonoporation: a multi-modal cell poration methodology through simultaneous application of electric field and ultrasonic wave
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DOI：
10.1039/c3lc40877a
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发表时间：
2013-01-01
期刊：
影响因子：
6.1
通讯作者：
Han, Arum
中科院分区：
文献类型：
--
作者：
Longsine-Parker, Whitney;Wang, Han;Han, Arum
本发明提供了一种微流体装置，其同时应用微电穿孔和微声穿孔所需的条件，以流通方案向哺乳动物细胞中高效率和高通量地递送分子。这种同时应用电场和超声波的多模式穿孔微器件是通过三维（3D）微电极方案实现的，其中电极既用作电穿孔电极又用作细胞流动通道，使得声波可以垂直于电场同时施加到流过微流体通道的细胞。这种3D微电极配置还允许施加均匀的电场，同时使该装置与荧光显微镜兼容。假设在垂直方向上同时施加两个不同的场（电场和声波）允许在降低的穿孔强度下沿细胞膜的两个轴沿着形成瞬时孔，因此最大化递送效率，同时最小化细胞死亡。微流控电声穿孔系统的特征在于将小分子递送到哺乳动物细胞中，并且显示平均穿孔效率为95.6%，细胞存活率为97.3%。该概念验证结果表明，与单独的电穿孔或声致穿孔相比，通过将电穿孔和声致穿孔组合在一起，可以实现分子递送效率的显著改善，同时保持高细胞活力。据我们所知，这里提出的微流体电声穿孔装置是第一个同时使用电场和超声波的多模式细胞穿孔装置。这种新的多模式细胞穿孔策略和系统预期在小分子治疗剂的递送中具有广泛的应用，并且最终在大分子递送中具有广泛的应用，例如基因转染应用，其中高递送效率和高活力是至关重要的。
A microfluidic device that simultaneously applies the conditions required for microelectroporation and microsonoporation in a flow-through scheme toward high-efficiency and high-throughput molecular delivery into mammalian cells is presented. This multi-modal poration microdevice using simultaneous application of electric field and ultrasonic wave was realized by a three-dimensional (3D) microelectrode scheme where the electrodes function as both electroporation electrodes and cell flow channel so that acoustic wave can be applied perpendicular to the electric field simultaneously to cells flowing through the microfluidic channel. This 3D microelectrode configuration also allows a uniform electric field to be applied while making the device compatible with fluorescent microscopy. It is hypothesized that the simultaneous application of two different fields (electric field and acoustic wave) in perpendicular directions allows formation of transient pores along two axes of the cell membrane at reduced poration intensities, hence maximizing the delivery efficiency while minimizing cell death. The microfluidic electro-sonoporation system was characterized by delivering small molecules into mammalian cells, and showed average poration efficiency of 95.6% and cell viability of 97.3%. This proof of concept result shows that by combining electroporation and sonoporation together, significant improvement in molecule delivery efficiency could be achieved while maintaining high cell viability compared to electroporation or sonoporation alone. The microfluidic electro-sonoporation device presented here is, to the best of our knowledge, the first multi-modal cell poration device using simultaneous application of electric field and ultrasonic wave. This new multi-modal cell poration strategy and system is expected to have broad applications in delivery of small molecule therapeutics and ultimately in large molecule delivery such as gene transfection applications where high delivery efficiency and high viability are crucial.