Coplanar film electrodes facilitate bovine nuclear transfer cloning
Coplanar film electrodes facilitate bovine nuclear transfer cloning
复制标题
共面薄膜电极促进牛核转移克隆
DOI：
10.1007/s10544-009-9302-z
复制
发表时间：
2009
影响因子：
2.8
通讯作者：
B. Oback
中科院分区：
文献类型：
--
作者：
A. Clow;P. Gaynor;B. Oback
芯片上的自动化实验室系统提供了更高的吞吐量和可重复性，但微电极的实现目前依赖于平行板电极的小型化，这是耗时和昂贵的制造。开放式电融合室的电场模型表明，宽间距（≥2mm）的共面膜电极与平行板电极的融合率相似，前提是电池的位置大致在电极之间的中间。这一假设是通过电融合试验的牛卵细胞供体细胞对用于核移植克隆。以平行板电极为对照进行对比实验。胚胎卵裂球、卵泡细胞、胎儿和成人成纤维细胞作为NT供体细胞共面融合率≥90%。对于胚胎和成体细胞类型，共面电极与平行板电极的融合率无显著差异。对于两种电极几何形状，在计算的电场强度为2.33 kV/cm时，与成纤维细胞的融合效率最高。共面电极需要比平行极板电极大π/2倍的电压，当耦合放置在电极中间时，才能达到等效的场强。
Automated lab on chip systems offer increased throughput and reproducibility, but the implementation of microelectrodes presently relies on miniaturisation of parallel plate electrodes that are time consuming and costly to fabricate. Electric field modelling of open electrofusion chambers suggested that widely spaced (≥2 mm) coplanar film electrodes should result in similar cell fusion rates as parallel plate electrodes provided the cell positioning was roughly midway between the electrodes. This hypothesis was investigated by electrofusion trials of bovine oocyte-donor cell couplets used in nuclear transfer (NT) cloning. Comparative experiments with reference parallel plate electrodes were conducted as controls. Coplanar fusion rates ≥ 90% were demonstrated for embryonic blastomeres, follicular cells and fetal and adult fibroblasts as NT donor cells. For embryonic and adult cell types, there was no significant difference in fusion rate between coplanar and parallel plate electrodes. For both electrode geometries, fusion efficiency with adult fibroblasts was highest at a calculated field strength of 2.33 kV/cm. The coplanar electrodes required a voltage π/2 times greater than parallel plate electrodes to achieve equivalent field strength when the couplets are placed midway between the electrodes.
影响因子：
2.8
作者：
Tresset, G;Takeuchi, S
通讯作者：
Takeuchi, S