A comparison between polymeric microsphere and bacterial vectors for macrophage P388D1 gene delivery
A comparison between polymeric microsphere and bacterial vectors for macrophage P388D1 gene delivery
复制标题
DOI：
10.1007/s11095-008-9563-x
复制
发表时间：
2008-05-01
影响因子：
3.7
通讯作者：
Pfeifer, Blaine A.
中科院分区：
文献类型：
--
作者：
Parsa, Saba;Wang, Yong;Pfeifer, Blaine A.
目的：本研究的目的是比较细菌和聚合物基因递送装置向鼠巨噬细胞P388 D1细胞系递送质粒DNA的能力。乳酸-羟基乙酸共聚物（PLGA）和聚将（β-氨基酯）聚合物配制成微球，该微球物理包埋编码萤火虫荧光素酶报告基因的质粒DNA;而将相同的质粒生物转化到工程化的大肠杆菌菌株中，以产生重组的嗜酸乳杆菌溶血素O。两个交付设备，然后测试基因交付和剂量的影响，使用巨噬细胞系与两种测定利用96孔高通量格式，以量化和比较每个载体type.Results：基因交付是可比的两种载体在较高的载体剂量，而较低的剂量显示出改进的微球载体的交付。递送效率（定义为荧光素酶测量值/mg细胞蛋白/递送的ng DNA）对于聚合物微球为881发光mg（-1）ng（-1），而对于细菌载体为171发光mg（-1）ng（-1）。第一个头对-聚合物和细菌基因递送载体之间的头部比较显示了聚合物微球的递送优势，病媒生产、储存和未来潜力。
Purpose: The purpose of this study was to compare bacterial and polymeric gene delivery devices for the ability to deliver plasmid DNA to a murine macrophage P388D1 cell line.Methods: An 85:15 ratio of poly(lactic-co-glycolic acid) (PLGA) and poly(beta-amino ester) polymers were formulated into microspheres that physically entrapped plasmid DNA encoding for the firefly luciferase reporter gene; whereas, the same plasmid was biologically transformed into a strain of Escherichia coli engineered to produce recombinant listeriolysin O. The two delivery devices were then tested for gene delivery and dosage effects using a macrophage cell line with both assays taking advantage of a 96-well high throughput format to quantify and compare each vector type.Results: Gene delivery was comparable for both vectors at higher vector dosages while lower dosages showed an improved delivery for the microsphere vectors. Delivery efficiency (defined as luciferase measurement/mg cellular protein/ng DNA delivered) was 881 luminescence mg(-1) ng(-1) for polymeric microspheres compared to 171 luminescence mg(-1) ng(-1) for the bacterial vectors.Conclusions: A first head-to-head comparison between polymeric and bacterial gene delivery vectors shows a delivery advantage for polymeric microspheres that must also be evaluated in light of vector production, storage, and future potential.