Phosphate-buffered saline-based nucleofection of primary endothelial cells.

Analytical Biochemistry
Jinjoo KangYoung-Kwon Hong

Abstract

Although various nonviral transfection methods are available, cell toxicity, low transfection efficiency, and high cost remain hurdles for in vitro gene delivery in cultured primary endothelial cells. Recently, unprecedented transfection efficiency for primary endothelial cells has been achieved due to the newly developed nucleofection technology that uses a combination of novel electroporation condition and specific buffer components that stabilize the cells in the electrical field. Despite superior transfection efficiency and cell viability, high cost of the technology has discouraged cardiovascular researchers from liberally adopting this new technology. Here we report that a phosphate-buffered saline (PBS)-based nucleofection method can be used for efficient gene delivery into primary endothelial cells and other types of cells. Comparative analyses of transfection efficiency and cell viability for primary arterial, venous, microvascular, and lymphatic endothelial cells were performed using PBS. Compared with the commercial buffers, PBS can support equally remarkable nucleofection efficiency to both primary and nonprimary cells. Moreover, PBS-mediated nucleofection of small interfering RNA (siRNA) showed more than 90% knockd...Continue Reading

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Citations

Jan 31, 2014·Journal of Neurochemistry·Gregory SaiaGrace Gill
Jun 8, 2015·European Neuropsychopharmacology : the Journal of the European College of Neuropsychopharmacology·Raquel PinachoBelén Ramos
Mar 10, 2016·Cartilage·Justin ParrenoRita Kandel
Nov 19, 2014·Molecular and Cellular Biology·Chen LiVijay K Kalra
Mar 3, 2010·The Journal of Immunology : Official Journal of the American Association of Immunologists·Caryn S Gonsalves, Vijay K Kalra
Oct 16, 2009·The Journal of Immunology : Official Journal of the American Association of Immunologists·Samantha M YeligarVijay K Kalra
Mar 1, 2020·BMC Molecular and Cell Biology·Iqbal HyderWilfried A Kues
Nov 23, 2011·Analytical Chemistry·Yongjing ChenPurnendu K Dasgupta
Jul 28, 2018·Chemical Reviews·Martin P StewartKlavs F Jensen

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