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Viromer RED和脂质体转染试剂在向神经胶质细胞瞬时导入基因中的比较效用

The Comparative Utility of Viromer RED and Lipofectamine for Transient Gene Introduction into Glial Cells.

作者信息

Rao Sudheendra, Morales Alejo A, Pearse Damien D

机构信息

The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136, USA.

The Miami Project to Cure Paralysis, University of Miami Miller School of Medicine, Miami, FL 33136, USA ; The Departments of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL 33136, USA ; The Neuroscience Program, University of Miami Miller School of Medicine, Miami, FL 33136, USA ; The Interdisciplinary Stem Cell Institute, University of Miami Miller School of Medicine, Miami, FL 33136, USA.

出版信息

Biomed Res Int. 2015;2015:458624. doi: 10.1155/2015/458624. Epub 2015 Oct 11.

Abstract

The introduction of genes into glial cells for mechanistic studies of cell function and as a therapeutic for gene delivery is an expanding field. Though viral vector based systems do exhibit good delivery efficiency and long-term production of the transgene, the need for transient gene expression, broad and rapid gene setup methodologies, and safety concerns regarding in vivo application still incentivize research into the use of nonviral gene delivery methods. In the current study, aviral gene delivery vectors based upon cationic lipid (Lipofectamine 3000) lipoplex or polyethylenimine (Viromer RED) polyplex technologies were examined in cell lines and primary glial cells for their transfection efficiencies, gene expression levels, and toxicity. The transfection efficiencies of polyplex and lipoplex agents were found to be comparable in a limited, yet similar, transfection setting, with or without serum across a number of cell types. However, differential effects on cell-specific transgene expression and reduced viability with cargo loaded polyplex were observed. Overall, our data suggests that polyplex technology could perform comparably to the market dominant lipoplex technology in transfecting various cells lines including glial cells but also stress a need for further refinement of polyplex reagents to minimize their effects on cell viability.

摘要

将基因导入神经胶质细胞以进行细胞功能的机制研究并作为基因递送的一种治疗方法,是一个不断发展的领域。尽管基于病毒载体的系统确实表现出良好的递送效率和转基因的长期表达,但对瞬时基因表达的需求、广泛而快速的基因设置方法以及对体内应用的安全性担忧,仍然促使人们对使用非病毒基因递送方法进行研究。在当前的研究中,基于阳离子脂质(Lipofectamine 3000)脂质体或聚乙烯亚胺(Viromer RED)多聚体技术的非病毒基因递送载体,在细胞系和原代神经胶质细胞中进行了转染效率、基因表达水平和毒性的检测。在有限但相似的转染条件下,无论有无血清,在多种细胞类型中,多聚体和脂质体试剂的转染效率相当。然而,观察到对细胞特异性转基因表达有不同影响,且负载了货物的多聚体导致细胞活力降低。总体而言,我们的数据表明,在转染包括神经胶质细胞在内的各种细胞系时,多聚体技术的表现可能与市场主导的脂质体技术相当,但也强调需要进一步优化多聚体试剂,以尽量减少它们对细胞活力的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a978/4619820/212873da241b/BMRI2015-458624.001.jpg

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