Using magnetic nanoparticles for gene transfer to neural stem cells: stem cell propagation method influences outcomes.
Using magnetic nanoparticles for gene transfer to neural stem cells: stem cell propagation method influences outcomes.
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DOI：
10.3390/jfb6020259
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发表时间：
2015-04-24
影响因子：
4.8
通讯作者：
Chari DM
中科院分区：
文献类型：
--
作者：
Pickard MR;Adams CF;Barraud P;Chari DM
基因工程神经干细胞（NSC）移植提供了一个关键的战略，以加强神经修复释放治疗生物分子到损伤部位。神经干细胞的遗传修饰在很大程度上依赖于病毒载体，但细胞毒性效应促使非病毒替代品的发展，如磁性纳米颗粒（MNP）。NSC在实验室中以3-D悬浮“神经球”或2-D粘附“单层”的形式繁殖。使用振荡磁场（“磁转染技术”）部署的MNP介导有效的基因转移到神经球，但这种方法对单层的功效是未知的。重要的是解决这个问题，因为振荡磁场显著增强了移植细胞中基于MNP的转染（例如，星形胶质细胞和少突胶质细胞前体）作为单层增殖。我们首次报告，振荡磁场增强MNP为基础的转染报告和功能（碱性成纤维细胞生长因子; FGF 2）基因在单层培养产生高转染与神经球。转染的神经干细胞表现出高活力，并可以重新形成神经球，这是很重要的，因为神经球产生更高的移植后活力比单层细胞。我们的研究结果表明，振荡磁场和单层形式的组合产生的MNP介导的基因转移到神经干细胞的最高效率，提供了一个可行的非病毒替代遗传修饰的这一重要的神经细胞移植人口。
Genetically engineered neural stem cell (NSC) transplants offer a key strategy to augment neural repair by releasing therapeutic biomolecules into injury sites. Genetic modification of NSCs is heavily reliant on viral vectors but cytotoxic effects have prompted development of non-viral alternatives, such as magnetic nanoparticle (MNPs). NSCs are propagated in laboratories as either 3-D suspension “neurospheres” or 2-D adherent “monolayers”. MNPs deployed with oscillating magnetic fields (“magnetofection technology”) mediate effective gene transfer to neurospheres but the efficacy of this approach for monolayers is unknown. It is important to address this issue as oscillating magnetic fields dramatically enhance MNP-based transfection in transplant cells (e.g., astrocytes and oligodendrocyte precursors) propagated as monolayers. We report for the first time that oscillating magnetic fields enhanced MNP-based transfection with reporter and functional (basic fibroblast growth factor; FGF2) genes in monolayer cultures yielding high transfection versus neurospheres. Transfected NSCs showed high viability and could re-form neurospheres, which is important as neurospheres yield higher post-transplantation viability versus monolayer cells. Our results demonstrate that the combination of oscillating magnetic fields and a monolayer format yields the highest efficacy for MNP-mediated gene transfer to NSCs, offering a viable non-viral alternative for genetic modification of this important neural cell transplant population.