Liver sinusoidal endothelial cells as possible vehicles for gene therapy: a comparison between plasmid-based and lentiviral gene transfer techniques.

Jesus Paez-Cortez, Ramon Montano, John Iacomini, Jose Cardier
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引用次数: 6

Abstract

Unlabelled: Liver sinusoidal endothelial cells (LSECs) constitute an attractive target for gene therapy of several liver and systemic diseases. However, there are few reports showing an efficient plasmid-based or viral methodology to deliver recombinant genes into these cells. In the present study, the authors evaluated in vitro gene transfer efficiency of standard plasmid-based techniques (i.e., electroporation, lipofection, and calcium phosphate) and lentiviral-mediated gene transduction into primary murine LSECs, using reporter genes. The results show that electroporation is the most effective in vitro plasmid-gene transfer method to deliver GFP into LSECs (31%), as compared with lipofection and calcium phosphate transfection (6% and 4%, respectively). However, lentiviral transduction resulted in higher, efficient, and stable gene transfer (70%) as compared with plasmid-based techniques.

Conclusions: The highly efficient gene expression obtained by lentiviral transduction and electroporation shows that these methodologies are highly reliable systems for gene transfer into LSECs.

肝窦内皮细胞作为基因治疗的可能载体:质粒和慢病毒基因转移技术的比较。
未标记:肝窦内皮细胞(LSECs)是几种肝脏和全身性疾病基因治疗的一个有吸引力的靶点。然而,很少有报道显示一种有效的基于质粒或病毒的方法将重组基因传递到这些细胞中。在本研究中,作者利用报告基因评估了标准质粒技术(即电穿孔、脂肪感染和磷酸钙)和慢病毒介导的基因转导到原代小鼠LSECs的体外基因转移效率。结果表明,与脂肪转染和磷酸钙转染(分别为6%和4%)相比,电穿孔是将GFP传递到LSECs的最有效的体外质粒基因转移方法(31%)。然而,与基于质粒的技术相比,慢病毒转导导致更高、高效和稳定的基因转移(70%)。结论:通过慢病毒转导和电穿孔获得的高效基因表达表明,这些方法是高度可靠的基因转移系统。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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