Cell internalization of magnetic nanoparticles using transfection agents
Cell internalization of magnetic nanoparticles using transfection agents
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DOI：
10.2310/7290.2006.00028
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发表时间：
2007-01-01
影响因子：
2.8
通讯作者：
Josephson, Lee
中科院分区：
文献类型：
--
作者：
Montet-Abou, Karin;Montet, Xavier;Josephson, Lee
用Feridex IV进行转染剂（TFA）诱导的磁性细胞标记是一种有吸引力的加载细胞的方法，因为它采用了氧化铁的药物来源。虽然有吸引力，但该方法有两个明显的缺点。首先，它需要混合带正电荷的转染剂和带负电荷的磁性纳米颗粒，并且所导致的纳米颗粒表面电荷的损失导致纳米颗粒沉淀。其次，它可以导致纳米颗粒吸附到细胞表面，而不是内化。菲立磁（和相关的葡聚糖）的内化是重要的，因为葡聚糖细胞外部可以与抗葡聚糖抗体（通常存在于人群中）反应，并引发抗体介导的细胞毒性。在这里，我们采用了三种测定法来选择菲立磁/TFA混合物，以最大限度地减少纳米颗粒沉淀和表面吸附：（1）沉淀或稳定性测定（光散射），（2）标记细胞测定（磁性过滤器保留的细胞百分比），和（3）基于抗葡聚糖的纳米颗粒内化测定。负载有菲立磁/鱼精蛋白的细胞具有内化的铁，而负载有菲立磁/Lipofectamine的细胞具有表面吸附的铁。最佳上样条件为10 μ g/Feridex和3 μ g/mL硫酸鱼精蛋白。需要仔细选择用菲立磁和TFA加载细胞的条件，以最大限度地减少纳米颗粒沉淀和葡聚糖吸附到细胞表面。
Transfection agent (TFA)-induced magnetic cell labeling with Feridex IV is an attractive method of loading cells because it employs a pharmaceutical source of iron oxide. Although attractive, the method has two significant drawbacks. First, it requires mixing positively charged transfection agents and negatively charged magnetic nanoparticles, and the resulting loss of nanoparticle surface charge causes nanoparticle precipitation. Second, it can result in nanoparticle adsorption to the cell surface rather than internalization. Internalization of Feridex (and associated dextran) is important since dextran cell exterior can react with the antidextran antibodies, commonly present in human populations, and trigger an antibody-mediated cytotoxicity. Here we employed three assays for selecting Feridex/TFA mixtures to minimize nanoparticle precipitation and surface adsorption: (1) an assay for precipitation or stability (light scattering), (2) an assay for labeled cells (percentage of cells retained by a magnetic filter), and (3) an antidextran-based assay for nanoparticle internalization. Cells loaded with Feridex/protamine had internalized iron, whereas cells loaded with Feridex/Lipofectamine had surf ace-adsorbed iron. Optimal conditions for loading cells were 10 mu g/Feridex and 3 mu g/mL protamine sulfate. Conditions for loading cells with Feridex and a TFA need to be carefully selected to minimize nanoparticle precipitation and dextran adsorption to the cell surface.