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纳米磁激活作为一种控制核酸递送效果的方法。

Nanomagnetic activation as a way to control the efficacy of nucleic acid delivery.

作者信息

Grześkowiak Bartosz F, Sánchez-Antequera Yolanda, Hammerschmid Edelburga, Döblinger Markus, Eberbeck Dietmar, Woźniak Anna, Słomski Ryszard, Plank Christian, Mykhaylyk Olga

机构信息

Institute of Experimental Oncology and Therapy Research, KLinikum rechts der Isar der Technischen Universität München, Ismaninger Str. 22, 81675, Munich, Germany.

出版信息

Pharm Res. 2015 Jan;32(1):103-21. doi: 10.1007/s11095-014-1448-6. Epub 2014 Jul 18.

Abstract

PURPOSE

To explore the potential of magnetofection in delivering pDNA to primary mouse embryonic fibroblasts (PMEFs) and porcine fetal fibroblasts (PFFs) and investigate an effect of magnetic cell labeling on transfection efficacy.

METHODS

The formulation and a dose of the magnetic vector were optimized. The efficacy of the procedure was quantified by vector internalization, transgene expression and cell iron loading upon specific labeling with Ab-conjugated magnetic beads or non-specific labeling with MNPs.

RESULTS

Up to sixty percent of PMEF and PFF cells were transfected at low pDNA doses of 4-16 pg pDNA/cell. Specific labeling of the PMEFs with MNPs, resulted in a 3- and 2-fold increase in pDNA internalization upon magnetofection and lipofection, respectively, that yielded a 2-4-fold increase in percent of transgene-expressing cells. Non-specific cell labeling had no effect on the efficacy of the reporter expression, despite the acquisition of similar magnetic moments per cell. In PFFs, specific magnetic labeling of the cell surface receptors inhibited internalization and transfection efficacy.

CONCLUSIONS

Magnetic labeling of cell-surface receptors combined with the application of an inhomogenous magnetic field (nanomagnetic activation) can affect the receptor-mediated internalization of delivery vectors and be used to control nucleic acid delivery to cells.

摘要

目的

探讨磁转染在将质粒DNA(pDNA)导入原代小鼠胚胎成纤维细胞(PMEF)和猪胎儿成纤维细胞(PFF)中的潜力,并研究磁性细胞标记对转染效率的影响。

方法

对磁性载体的配方和剂量进行优化。通过载体内化、转基因表达以及用抗体偶联磁珠进行特异性标记或用磁性纳米颗粒(MNP)进行非特异性标记后的细胞铁负载来量化该方法的效率。

结果

在低至4 - 16 pg pDNA/细胞的pDNA剂量下,高达60%的PMEF和PFF细胞被转染。用MNP对PMEF进行特异性标记,分别使磁转染和脂质体转染时的pDNA内化增加了3倍和2倍,这使得转基因表达细胞的百分比增加了2 - 4倍。非特异性细胞标记对报告基因表达效率没有影响,尽管每个细胞获得了相似的磁矩。在PFF中,细胞表面受体的特异性磁性标记抑制了内化和转染效率。

结论

细胞表面受体的磁性标记与不均匀磁场的应用(纳米磁激活)相结合,可以影响递送载体的受体介导内化,并可用于控制核酸向细胞的递送。

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