Gene delivery through cell culture substrate adsorbed DNA complexes
Gene delivery through cell culture substrate adsorbed DNA complexes
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DOI：
10.1002/bit.20393
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发表时间：
2005-05-05
影响因子：
3.8
通讯作者：
Shea, LD
中科院分区：
文献类型：
--
作者：
Bengali, Z;Pannier, AK;Shea, LD
有效的基因传递是生物技术的一个基本目标，在基础科学和应用科学中有许多应用。底物介导的递送和反向转染通过在细胞接种之前将质粒固定到细胞培养底物上来增加细胞微环境中DNA的浓度，从而增强基因转移。在这份报告中，我们研究了基因传递的质粒与阳离子聚合物（复合物）或脂质（脂复合物），随后固定到细胞培养或生物材料基板的吸附。聚合复合物和脂质复合物被吸附到组织培养聚苯乙烯或血清吸附的组织培养聚苯乙烯上，固定的DNA量随暴露时间的增加而增加，沉积速率和最终沉积量取决于基质和复合物的性质。对于复合物，血清修饰增强报告基因的表达高达1 500倍，相对于未修饰的基板，并产生相当于或更大的表达相比，团注输送。对于lipoplexes，血清修饰显着增加了转染细胞的数量相对于未修饰的基板，但提供了类似的表达水平。相对于推注递送，固定化复合物以改善的细胞活力捕获原代细胞。最后，这种基质介导的递送方法被扩展到广泛使用的生物材料，聚（丙交酯-共-乙交酯）。将DNA复合物固定到组织培养聚苯乙烯基质上可以是用于增强体外研究的基因递送的有用工具。此外，使该系统适应生物材料可以促进应用于诸如组织工程的领域。(c)2005 Wiley Periodicals，Inc.
Efficient gene delivery is a fundamental goal of biotechnology and has numerous applications in both basic and applied science. Substrate-mediated delivery and reverse transfection enhance gene transfer by increasing the concentration of DNA in the cellular microenvironment through immobilizing a plasmid to a cell culture substrate prior to cell seeding. In this report, we examine gene delivery of plasmids that were complexed with cationic polymers (polyplexes) or lipids (lipoplexes) and subsequently immobilized to cell culture or biomaterial substrates by adsorption. Polyplexes and lipoplexes were adsorbed to either tissue culture polystyrene or serum-adsorbed tissue culture polystyrene, The quantity of DNA immobilized increased with time of exposure, and the deposition rate and final amount deposited depended upon the properties of the substrate and complex. For polyplexes, serum modification enhanced reporter gene expression up to 1 500-fold relative to unmodified substrates and yielded equivalent or greater expression compared to bolus delivery. For lipoplexes, serum modification significantly increased the number of transfected cells relative to unmodified substrates yet provided similar levels of expression. Immobilized complexes transfect primary cells with improved cellular viability relative to bolus delivery. Finally, this substrate-mediated delivery approach was extended to a widely used biomaterial, poly(lactide-co-glycolide). Immobilization of DNA complexes to tissue culture polystyrene substrates can be a useful tool for enhancing gene delivery for in vitro studies. Additionally, adapting this system to biornaterials may facilitate application to fields such as tissue engineering. (c) 2005 Wiley Periodicals, Inc.