Micronozzle Array Enhanced Sandwich Electroporation of Embryonic Stem Cells
Micronozzle Array Enhanced Sandwich Electroporation of Embryonic Stem Cells
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DOI：
10.1021/ac902041h
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发表时间：
2010-01-01
影响因子：
7.4
通讯作者：
Lee, L. James
中科院分区：
文献类型：
--
作者：
Fei, Zhengzheng;Hu, Xin;Lee, L. James
电穿孔是胚胎干细胞转染最流行的非病毒基因转移方法之一。然而，批量电穿孔技术需要高电场，并且在随机分布的细胞之间提供不均匀的电场分布，导致转染效率和细胞活力有限，特别是对于少量细胞而言。我们在这里介绍一种使用明确的微喷嘴阵列的膜夹心电穿孔系统。该装置能够以良好的性能转染数百至数百万个细胞。处理少量细胞（即一百个）的能力为使用难以收获的患者细胞进行药物动力学研究提供了巨大的潜力。初始跨膜电位分布和碘化丙啶（PI）染料扩散实验的数值模拟证明了微喷嘴阵列提供的高度聚焦和局部电场强度相对于传统批量电穿孔的优势。
Electroporation is one of the most popular nonviral gene transfer methods for embryonic stem cell transfection. Bulk electroporation techniques, however, require a high electrical field and provide a nonuniform electrical field distribution among randomly distributed cells, leading to limited transfection efficiency and cell viability, especially for a low number of cells. We present here a membrane sandwich electroporation system using a well-defined micronozzle array. This device is capable of transfecting hundred to millions of cells with good performance. The ability to treat a small number of cells (i.e., a hundred) offers great potential to work with hard-to-harvest patient cells for pharmaceutical kinetic studies. Numerical simulation of the initial transmembrane potential distribution and propidium iodide (PI) dye diffusion experiments demonstrated the advantage of highly focused and localized electric field strength provided by the micronozzle array over conventional bulk electroporation.