Static magnetic field reduced exogenous oligonucleotide uptake by spermatozoa using magnetic nanoparticle gene delivery system
Static magnetic field reduced exogenous oligonucleotide uptake by spermatozoa using magnetic nanoparticle gene delivery system
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DOI：
10.1016/j.jmmm.2015.11.057
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发表时间：
2016-03
影响因子：
2.7
通讯作者：
Samira Katebi;A. Esmaeili;K. Ghaedi
中科院分区：
文献类型：
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作者：
Samira Katebi;A. Esmaeili;K. Ghaedi
精子可以将外源寡核苷酸引入卵母细胞。精子介导的基因转移(SMGT)效率低的最重要原因是精子对外源DNA的摄取率低。本研究的目的是用磁流变法评价静磁场对精子摄取外源寡核苷酸的影响。与标记的寡核苷酸结合的磁性纳米颗粒(MNPs)用于提高公鸡精子摄取外源寡核苷酸的效率。我们使用高场/高梯度磁铁(NdFeB)来促进和加速外源DNA在精子表面的沉积。用流式细胞仪分析精子的存活率和摄取外源寡核苷酸的百分比。流式细胞仪分析显示，在外源寡核苷酸溶液和MNPs中孵育时，公鸡精子对外源寡核苷酸的摄取显著增加(P<；0.001)。然而，在磁偏析过程中施加静磁场，外源寡核苷酸摄取量显著减少(P<；0.05)。本研究的结果表明，MNPs能有效地提高鸡精子对外源寡核苷酸的摄取，但与其他研究不同的是，静磁场不仅不能有效地提高公鸡精子对外源寡核苷酸的摄取，而且还导致磁性纳米粒在基因转移中的效率降低。
Spermatozoa could introduce exogenous oligonucleotides of interest to the oocyte. The most important reason of low efficiency of sperm mediated gene transfer (SMGT) is low uptake of exogenous DNA by spermatozoa. The aim of this study was to evaluate the effects of static magnetic field on exogenous oligonucleotide uptake of spermatozoa using magnetofection method. Magnetic nanoparticles (MNPs) associated with the labeled oligonucleotides were used to increase the efficiency of exogenous oligonucleotide uptake by rooster spermatozoa. We used high-field/high-gradient magnet (NdFeB) to enhance and accelerate exogenous DNA sedimentation at the spermatozoa surface. Flow cytometry analysis was performed to measure viability and percentage of exogenous oligonucleotide uptake by sperm. Flow cytometry analysis showed a significant increase in exogenous oligonucleotide uptake by rooster spermatozoa (P<0.001) when spermatozoa were incubated in exogenous oligonucleotide solution and MNPs. However, by applying static magnetic field during magnetofection method, a significant decrease in exogenous oligonucleotide uptake was observed (P<0.05). Findings of this study showed that MNPs were effective to increase exogenous oligonucleotide uptake by rooster spermatozoa; however unlike others studies, static magnetic field, was not only ineffective to enhance exogenous oligonucleotide uptake by rooster spermatozoa but also led to reduction in efficiency of magnetic nanoparticles in gene transfer.