Lentiviral vector-mediated genetic modification of human neural progenitor cells for ex vivo gene therapy
Lentiviral vector-mediated genetic modification of human neural progenitor cells for ex vivo gene therapy
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DOI：
10.1016/j.jneumeth.2007.02.022
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发表时间：
2007-07-30
影响因子：
3
通讯作者：
Svendsen, Clive N.
中科院分区：
文献类型：
--
作者：
Capowski, Elizabeth E.;Schneider, Bernard L.;Svendsen, Clive N.
人神经祖细胞（hNPC）作为一种将治疗性蛋白质递送至中枢神经系统的离体系统具有巨大的潜力。当作为聚集体（称为神经球）培养时，hNPC能够在体外显著扩增。在目前的研究中，我们提出了一种稳健的方法，慢病毒载体介导的基因传递到hNPC，保持神经球培养的分化和增殖特性，同时尽量减少病毒载体的使用量，并控制每个群体的插入位点的数量。即使在hNPC分化为神经元和星形胶质细胞后，这种方法也能实现长期稳定的表达，并允许产生等效的转基因hNPC群体。此外，体外分析预测了转基因株系移植到帕金森病啮齿动物模型中时的体内行为。提出的方法提供了一个强大的工具，用于评估因素，如启动子系统或不同的转基因对这些细胞的治疗效用的影响。(c)2007 Elsevier B.V.保留所有权利。
Human neural progenitor cells (hNPC) hold great potential as an ex vivo system for delivery of therapeutic proteins to the central nervous system. When cultured as aggregates, termed neurospheres, hNPC are capable of significant in vitro expansion. In the current study, we present a robust method for lentiviral vector-mediated gene delivery into hNPC that maintains the differentiation and proliferative properties of neurosphere cultures while minimizing the amount of viral vector used and controlling the number of insertion sites per population. This method results in long-term, stable expression even after differentiation of the hNPC to neurons and astrocytes and allows for generation of equivalent transgenic, populations of hNPC. In addition, the in vitro analysis presented predicts the behavior of transgenic lines in vivo when transplanted into a rodent model of Parkinson's disease. The methods presented provide a powerful tool for assessing the impact of factors such as promoter systems or different transgenes on the therapeutic utility of these cells. (c) 2007 Elsevier B.V. All rights reserved.