Mesenchymal stem cells, MG63 and HEK293 transfection using chitosan-DNA nanoparticles
Mesenchymal stem cells, MG63 and HEK293 transfection using chitosan-DNA nanoparticles
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DOI：
10.1016/s0142-9612(02)00507-0
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发表时间：
2003-03-01
期刊：
影响因子：
14
通讯作者：
Fernandes, JC
中科院分区：
文献类型：
--
作者：
Corsi, K;Chellat, F;Fernandes, JC
将阳离子聚合物与β-半乳糖DNA质粒络合，合成了壳聚糖-DNA纳米粒，以研究壳聚糖开发非病毒基因递送系统的效果，该系统可优化用于有效的基因治疗。用荧光胺和PicoGreen法确定了最佳结合条件。用电子显微镜观察了DNA在纳米颗粒中的分布，用原子力显微镜分析了纳米颗粒的大小和形态。首次对人骨髓间充质干细胞(MSCs)、人骨肉瘤细胞(MG63)和人胚胎肾细胞(HEK293)进行了转染性评价。用LipofetAMINE(TM)2000(LF)试剂作对照。用3-[4，5-二甲基噻唑-2-基]-2，5-二苯基四氮唑(四甲基偶氮唑蓝)比色法观察壳聚糖DNA纳米粒对细胞活力的影响。形成的纳米颗粒直径小于100 nm，DNA分布均匀。HEK293细胞的转染率优于MG63细胞和MSCs，但与超过50%的LF毒性相比，复合体的细胞毒性最小。这些结果表明，壳聚糖-DNA纳米粒具有良好的非病毒基因传递特性，具有细胞类型依赖性和无细胞毒性。(C)2002爱思唯尔科学有限公司。保留所有权利。
Chitosan-DNA nanoparticles were synthesized from the complexation of the cationic polymer with a beta-gal DNA plasmid, in order to study the efficacy of chitosan to develop a non-viral gene delivery system that can be optimized for efficient gene therapy. The optimal binding conditions were determined with the fluorescamine and PicoGreen assays. DNA distribution within the nanoparticle was visualized by electron transmission microscopy, while the size and morphology were assessed by atomic force microscopy. The transfection potential was evaluated for the first time on human mesenchymal stem cells (MSCs), on human osteosarcoma cells (MG63) and on human embryonic kidney cells (HEK293). The LipofectAMINE(Tm) 2000 (LF) reagent was used in comparison. The effect of chitosan-DNA nanoparticles on cell viability was illustrated with the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. The nanoparticles formed are of a diameter inferior to 100 nm with a homogenous distribution of DNA. The transfection of HEK293 cells is superior to that seen with MG63 cells and MSCs, however not surpassing that seen with LF Minimal cytotoxicity is seen with the polyplexes compared to greater than 50% toxicity with LF. These results suggest that chitosan-DNA nanoparticles have favorable characteristics for non-viral gene delivery, are cell type dependent and not cytotoxic. (C) 2002 Elsevier Science Ltd. All rights reserved.