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首页> 外文期刊>Human gene therapy. Clinical development >Direct Head-To-Head Comparison of Cationic Liposome-Mediated Gene Delivery to Mesenchymal Stem/Stromal Cells of Different Human Sources: A Comprehensive Study
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Direct Head-To-Head Comparison of Cationic Liposome-Mediated Gene Delivery to Mesenchymal Stem/Stromal Cells of Different Human Sources: A Comprehensive Study

机译：阳离子脂质体介导的基因递送到不同人类来源的间充质干细胞/间质细胞的直接头对头比较：一项全面的研究

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Nonviral gene delivery to human mesenchymal stem/stromal cells (MSC) can be considered a very promising strategy to improve their intrinsic features, amplifying the therapeutic potential of these cells for clinical applications. In this work, we performed a comprehensive comparison of liposome-mediated gene transfer efficiencies to MSC derived from different human sources-bone marrow (BM MSC), adipose tissue-derived cells (ASC), and umbilical cord matrix (UCM MSC). The results obtained using a green fluorescent protein (GFP)-encoding plasmid indicated that MSC isolated from BM and UCM are more amenable to genetic modification when compared to ASC as they exhibited superior levels of viable, GFP+ cells 48 hr post-transfection, 58 (+-)7.1% and 54 (+-) 3.8%, respectively, versus 33 (+-)4.7%. For all cell sources, high cell recoveries (approx =50%) and viabilities (>85%) were achieved, and the transgene expression was maintained for 10 days. Levels of plasmid DNA uptake, as well as kinetics of transgene expression and cellular division, were also determined. Importantly, modified cells were found to retain their characteristic immunophenotypic profile and multilineage differentiation capacity. By using the lipofection protocol optimized herein, we were able to maximize transfection efficiencies to human MSC (maximum of 74% total GFP+ cells) and show that lipofection is a promising transfection strategy for MSC genetic modification, especially when a transient expression of a therapeutic gene is required. Importantly, we also clearly demonstrated that intrinsic features of MSC from different sources should be taken into consideration when developing and optimizing strategies for MSC engineering with a therapeutic gene.
机译：将非病毒基因递送至人间充质干/基质细胞（MSC）被认为是改善其固有特征，扩大这些细胞在临床应用中的治疗潜力的非常有前途的策略。在这项工作中，我们对脂质体介导的基因转移效率进行了比较，该效率来自不同的人类来源-骨髓（BM MSC），脂肪组织衍生细胞（ASC）和脐带基质（UCM MSC）。使用绿色荧光蛋白（GFP）编码质粒获得的结果表明，与ASC相比，从BM和UCM分离的MSC更易于进行基因修饰，因为它们在转染后48小时显示出较高水平的可行GFP +细胞，58（ +-）7.1％和54（+-）3.8％，而33（+-）4.7％。对于所有细胞来源，均实现了高细胞回收率（约= 50％）和活力（> 85％），并且转基因表达保持了10天。还测定了质粒DNA的摄取水平，以及转基因表达和细胞分裂的动力学。重要的是，发现修饰的细胞保留其特征性免疫表型特征和多系分化能力。通过使用本文优化的脂转染方案，我们能够最大化对人MSC的转染效率（最大占总GFP +细胞的74％），并表明脂转染是用于MSC基因修饰的有前途的转染策略，尤其是当治疗性基因的瞬时表达时是必须的。重要的是，我们还清楚地表明，在开发和优化具有治疗基因的MSC工程的策略时，应考虑来自不同来源的MSC的固有特征。

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