Functionalization of microparticles with mineral coatings enhances non-viral transfection of primary human cells.
Functionalization of microparticles with mineral coatings enhances non-viral transfection of primary human cells.
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DOI：
10.1038/s41598-017-14153-x
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发表时间：
2017-10-27
影响因子：
4.6
通讯作者：
Murphy WL
中科院分区：
文献类型：
--
作者：
Khalil AS;Yu X;Xie AW;Fontana G;Umhoefer JM;Johnson HJ;Hookway TA;McDevitt TC;Murphy WL
基因传递到原代人类细胞是一项对生命科学研究和治疗应用至关重要的技术。然而，低效率的基因转移和不理想的安全性仍然是该技术发展的主要限制。在这里，我们描述了一种基于材料的方法，即生物可吸收矿物涂层的应用改善了质粒DNA脂质体在几种主要人类细胞类型中的微颗粒转染。在这些矿物质包被的微颗粒（mcm）存在下，我们观察到转染效率提高了4倍，同时细胞毒性降低。我们确定了mcm改善转染的机制，以及在三维细胞结构中改善转染的涂层成分。该方法为二维和三维转染策略提供了有效的人原代成纤维细胞、间充质细胞和胚胎干细胞转染。这种基于mcm的转染是基因传递技术的一项进步，因为它代表了一种非病毒方法，可以实现高效、局部转染，并允许转染三维细胞结构。
Gene delivery to primary human cells is a technology of critical interest to both life science research and therapeutic applications. However, poor efficiencies in gene transfer and undesirable safety profiles remain key limitations in advancing this technology. Here, we describe a materials-based approach whereby application of a bioresorbable mineral coating improves microparticle-based transfection of plasmid DNA lipoplexes in several primary human cell types. In the presence of these mineral-coated microparticles (MCMs), we observed up to 4-fold increases in transfection efficiency with simultaneous reductions in cytotoxicity. We identified mechanisms by which MCMs improve transfection, as well as coating compositions that improve transfection in three-dimensional cell constructs. The approach afforded efficient transfection in primary human fibroblasts as well as mesenchymal and embryonic stem cells for both two- and three-dimensional transfection strategies. This MCM-based transfection is an advancement in gene delivery technology, as it represents a non-viral approach that enables highly efficient, localized transfection and allows for transfection of three-dimensional cell constructs.
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