Transfection of Cardiac Cells by Means of Laser-Assisted Optoporation

The development of tissue and genetic engineering approaches to cardiovascular therapy is hampered by our limited ability to manipulate gene expression in 3D tissue structures with cellular resolution. An important unmet need is heterogeneous gene expression in cardiac cells and tissues, which requires targeted delivery of foreign DNA into selected cells or regions. Several techniques have been used for targeted transfection, such as direct microinjection into cells or targeted electroporation. However, these techniques have limited throughput or spatial resolution of transfection. We developed a method for transfection of cardiac cells by means of laser-assisted optoporation and validated it using a standard confocal microscope. This technique allows for the transfection of selected cell types in the presence of other cell types provided that they are detectable with a microscope. This technique can work as a “gene printer” creating arbitrarily shaped areas of transfected cells. Keywords: laserporation; confocal fluorescent microscopy; targeted gene delivery; cardiac myocyte; transfection; optoporation; laser; cardiac myocytes; fibroblasts

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