A novel electroporation method using a capillary and wire-type electrode
A novel electroporation method using a capillary and wire-type electrode
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DOI：
10.1016/j.bios.2007.12.009
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发表时间：
2008-04-01
影响因子：
12.6
通讯作者：
Chang, Jun Keun
中科院分区：
文献类型：
--
作者：
Kim, Jeong Ah;Cho, Keunchang;Chang, Jun Keun
电穿孔被广泛用于实现基因转染。电穿孔中的一个常见问题是它比任何其他转染方法具有更低的活力。在这项研究中，我们开发了一种新的电穿孔装置，使用毛细管尖端和移液管，是有效的对广泛的哺乳动物细胞，包括细胞系，原代细胞和干细胞。毛细管电穿孔系统由于其具有小的表面积的线型电极而大大减少了电穿孔过程中的细胞死亡。实验结果还表明，细胞活力依赖于在电穿孔过程中由电解引起的pH的变化。此外，使用长而窄的毛细管与简单的移液相结合，缩短了电穿孔过程的总时间长达15分钟，即使在不同的条件下，24个样品。这些结果通过与常规电穿孔系统的比较得到支持。通过使用毛细管系统提高了转染率和细胞活力，其在一般细胞系如HeLa和COS-7中具有80%以上的高转染率，在难以转染的细胞如干细胞或原代细胞中具有50%以上的高转染率。本研究中使用的所有细胞类型的活力约为70-80%。(C)2007 Elsevier B. V.保留所有权利。
Electroporation is widely used to achieve gene transfection. A common problem in electroporation is that it has a lower viability than any other transfection method. In this study, we developed a novel electroporation device using a capillary tip and a pipette that was effective on a wide range of mammalian cells, including cell lines, primary cells, and stem cells. The capillary electroporation system considerably reduced cell death during electroporation because of its wire-type electrode, which has a small surface area. The experimental results also indicated that the cell viability was dependent on the change in pH induced by electrolysis during electroporation. Additionally, the use of a long and narrow capillary tube combined with simple pipetting shortened the overall time of the electroporation process by up to 15 min, even under different conditions with 24 samples. These results were supported by comparison with a conventional electroporation system. The transfection rate and the cell viability were enhanced by the use of the capillary system, which had a high transfection rate of more than 80% in general cell lines such as HeLa and COS-7, and more than 50% in hard-to-transfect cells such as stem or primary cells. The viability was approximately 70-80% in all cell types used in this study. (C) 2007 Elsevier B.V. All rights reserved.