Magnetic Nanoparticle-Mediated Gene Delivery to Two- and Three-Dimensional Neural Stem Cell Cultures: Magnet-Assisted Transfection and Multifection Approaches to Enhance Outcomes.
Magnetic Nanoparticle-Mediated Gene Delivery to Two- and Three-Dimensional Neural Stem Cell Cultures: Magnet-Assisted Transfection and Multifection Approaches to Enhance Outcomes.
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DOI：
10.1002/cpsc.23
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发表时间：
2017-02
影响因子：
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通讯作者：
M. Pickard;C. Adams;D. Chari
中科院分区：
文献类型：
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作者：
M. Pickard;C. Adams;D. Chari
神经干细胞在神经修复的移植治疗中具有很高的转化潜力。通过基因工程增强其治疗能力是再生神经学的一个重要目标。磁性纳米颗粒（MNP）是用于神经干细胞安全生物工程的主要非病毒载体，与病毒载体相比具有关键的翻译优势，包括安全性，可扩展性和易用性。本单元描述了悬浮液（神经球）和贴壁（单层）小鼠神经干细胞培养的生产方案。神经干细胞与MNP的基因工程和应用“磁转染”（磁场）或“多重转染”（重复转染）的方法，以提高基因传递。单层培养的磁转染实现了最佳转染，但神经球提供了移植后神经移植物存活的关键优势。一个协议，它允许每种方法的优点被组合成一个单一的程序移植。这些协议的适应其他MNP制剂被认为是，重点是程序的安全性评价。John Wiley & Sons，Inc.
Neural stem cells (NSCs) have high translational potential in transplantation therapies for neural repair. Enhancement of their therapeutic capacity by genetic engineering is an important goal for regenerative neurology. Magnetic nanoparticles (MNPs) are major non-viral vectors for safe bioengineering of NSCs, offering critical translational benefits over viral vectors, including safety, scalability, and ease of use. This unit describes protocols for the production of suspension (neurosphere) and adherent (monolayer) murine NSC cultures. Genetic engineering of NSCs with MNPs and the application of 'magnetofection' (magnetic fields) or 'multifection' (repeat transfection) approaches to enhance gene delivery are described. Magnetofection of monolayer cultures achieves optimal transfection, but neurospheres offer key advantages for neural graft survival post-transplantation. A protocol is presented which allows the advantageous features of each approach to be combined into a single procedure for transplantation. The adaptation of these protocols for other MNP preparations is considered, with emphasis on the evaluation of procedural safety. © 2017 by John Wiley & Sons, Inc.