Co-transfection with cDNA encoding the Bcl family of anti-apoptotic proteins improves the efficiency of transfection in primary fetal neural stem cells
Co-transfection with cDNA encoding the Bcl family of anti-apoptotic proteins improves the efficiency of transfection in primary fetal neural stem cells
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DOI：
10.1016/s0165-0270(02)00090-0
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发表时间：
2002-06
影响因子：
3
通讯作者：
You-Chan Kim;J. Shim;Y. Oh;H. Son;Yong-Sung Lee;Sang-Hun Lee
中科院分区：
文献类型：
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作者：
You-Chan Kim;J. Shim;Y. Oh;H. Son;Yong-Sung Lee;Sang-Hun Lee
原代神经干细胞转染效率低下是基因表达研究的主要障碍，旨在确定脑发育的机制。在我们最初发现脂质体介导的转染程序对神经干细胞具有相当大的毒性之后，我们进一步研究了是否可以通过防止细胞发生凋亡来提高转染效率。通过用原型抗凋亡基因（如bcl-2和bcl-xL）共转染细胞并用B27补充剂补充培养基，转染效率显著增强。抗凋亡基因共转染和B27补充剂的这种组合导致原代神经干细胞的约5%的转染效率，而对照转染中的转染效率小于0.2%。因此，这种方法和其他类似的方法，用于提高效率的体外培养神经干细胞可能有助于了解神经干细胞的自我更新及其分化成各种细胞谱系的机制。
Inefficiency in primary neural stem cell transfection is a major obstacle in gene expression research aimed at determining the mechanisms underlying brain development. Following our initial finding that liposome-mediated transfection procedures are fairly toxic to neural stem cells, we further examined whether transfection efficiency could be improved by preventing cells from undergoing apoptosis. Transfection efficiencies were markedly enhanced by co-transfection of cells with prototypic anti-apoptotic genes, such as bcl-2 and bcl-xL, and supplementing the culture medium with B27 Supplement. This combination of anti-apoptotic gene co-transfection and B27 Supplement resulted in approximately 5% transfection efficiency of primary neural stem cells, compared to less than 0.2% in control transfections. Therefore, this procedure and other similar approaches employed to enhance the efficiency of transfecting neural stem cells may facilitate the understanding of mechanisms underlying self-renewal of neural stem cells and their differentiation into various cell lineages.