Transfection agent induced nanoparticle cell loading.
Transfection agent induced nanoparticle cell loading.
复制标题
转染剂诱导纳米颗粒细胞负载。
DOI：
10.1162/15353500200505100
复制
发表时间：
2005
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;Weissleder,Ralph;Josephson,Lee
用磁性纳米粒子装载细胞，并通过高分辨率 MRI 跟踪它们在体内的命运，是增强细胞疗法（包括利用造血干细胞、神经祖细胞和 T 细胞的疗法）疗效的一种有吸引力的方法。使用 Feridex IV® 纳米颗粒辅助装载转染剂（内化剂）是一种颇具吸引力的装载方法，因为材料成本低廉，并且最终临床使用的监管障碍可能较低。因此，我们探索了 Feridex IV® 与三种内化剂鱼精蛋白 (PRO)、聚赖氨酸 (PLL) 和 lipofectamine (LFA) 之间的相互作用。 Feridex与内化剂反应形成聚集体，除非内化剂或Feridex大量过量存在。当 Jurkat T 细胞与 Feridex/LFA 或 Feridex/PRO 混合物一起孵育并通过离心洗涤时，纳米颗粒聚集体与细胞共纯化。对于C17.2细胞，大的氧化铁颗粒粘附在细胞表面。在 30 μg/mL Feridex 和 3 μg/mL LFA 下，内化主要由 LFA 介导，并且主要在细胞质中。然而，我们发现用Feridex和转染剂标记细胞的条件需要仔细选择，以避免表面吸附和纳米颗粒沉淀的问题。
Loading cells with magnetic nanoparticles, and tracking their fate in vivo by high resolution MRI, is an attractive approach for enhancing the efficacy of cell-based therapies including those utilizing hematopoietic stem cells, neuroprogenitor cells, and T cells. The transfection agent (internalization agent) assisted loading with the Feridex IV®nanoparticle is an attractive method of loading because of the low cost of materials, and possible low regulatory barriers for eventual clinical use. We therefore explored the interaction between Feridex IV®and three internalization agents protamine (PRO), polylysine (PLL), and lipofectamine (LFA). Feridex reacted with internalization agents to form aggregates, except when either the internalization agent or Feridex was present in large excess. When Jurkat T cells were incubated with Feridex/LFA or Feridex/PRO mixtures, and washed by centrifugation, nanoparticle aggregates co-purified with cells. With C17.2 cells large iron oxide particles adhered to the cell surface. At 30 μg/mL Feridex and 3 μg/mL LFA, internalization was largely mediated by LFA and was largely cytoplasmic. However, we found that the conditions used to label cells with Feridex and transfection agents need to be carefully selected to avoid the problems of surface adsorption and nanoparticle precipitation.