Department of Biomedical Sciences, Section of Physiology, University of Padua, via F. Marzolo 3, 35131, Padua, Italy.
Padua Neuroscience Center, University of Padua, via Orus 2/B, 35131, Padua, Italy.
Sci Rep. 2021 Jul 23;11(1):15126. doi: 10.1038/s41598-021-94620-8.
Electroporation is a widely used non-viral technique for the delivery of molecules, including nucleic acids, into cells. Recently, electronic microsystems that miniaturize the electroporation machinery have been developed as a new tool for genetic manipulation of cells in vitro, by integrating metal microelectrodes in the culture substrate and enabling electroporation in-situ. We report that non-faradic SiO thin film-insulated microelectrodes can be used for reliable and spatially selective in-situ electroporation of mammalian cells. CHO-K1 and SH-SY5Y cell lines and primary neuronal cultures were electroporated by application of short and low amplitude voltage transients leading to cell electroporation by capacitive currents. We demonstrate reliable delivery of DNA plasmids and exogenous gene expression, accompanied by high spatial selectivity and cell viability, even with differentiated neurons. Finally, we show that SiO thin film-insulated microelectrodes support a double and serial transfection of the targeted cells.
电穿孔是一种广泛应用的非病毒技术,用于将分子(包括核酸)递送入细胞。最近,电子微系统将电穿孔机械微型化,作为体外细胞遗传操作的新工具,通过在培养基板中集成金属微电极,并实现原位电穿孔。我们报告称,非法拉第的 SiO 薄膜绝缘微电极可用于可靠且空间选择性的原位哺乳动物细胞电穿孔。通过施加短而低幅度的电压瞬变,CHO-K1 和 SH-SY5Y 细胞系和原代神经元培养物发生电穿孔,从而通过电容电流导致细胞电穿孔。我们证明了 DNA 质粒和外源基因表达的可靠传递,同时具有高空间选择性和细胞活力,即使是分化的神经元也是如此。最后,我们表明 SiO 薄膜绝缘微电极支持靶向细胞的双重和串联转染。