Differential dielectric responses of chondrocyte and Jurkat cells in electromanipulation buffers.
Differential dielectric responses of chondrocyte and Jurkat cells in electromanipulation buffers.
复制标题
DOI：
10.1002/elps.201500119
复制
发表时间：
2015-07
期刊：
影响因子：
2.9
通讯作者：
Beskok A
中科院分区：
文献类型：
--
作者：
Sabuncu AC;Asmar AJ;Stacey MW;Beskok A
细胞电操作作为一种无标记的细胞操作和表征工具，最近获得了特别的兴趣。然而，电操纵，特别是介电泳（DEP），生物细胞的适用性是有限的细胞悬浮在缓冲液中含有较低的盐量相对于生理缓冲液。人们可能会质疑使用低电导率缓冲液（LCB）进行DEP分离，因为细胞在缺乏生理水平盐的缓冲液中受到应激。在LCB中，细胞泄漏离子并进行体积调节。因此，细胞在LCB中表现出时间依赖性DEP响应。在这项工作中，LCB中的细胞变化通过介电谱、细胞活力测定以及软骨细胞和Jurkats的基因表达来评估。结果表明，离子从细胞中泄漏，细胞质电导率，膜电容和电导增加。使用细胞介电数据计算两种细胞类型的可分离性因子，其定义DEP细胞分离的最佳条件。最佳的DEP分离条件随着LCB中细胞介电性质的演变而变化。遗传分析表明，悬浮在LCB中的软骨细胞的离子通道蛋白的表达没有变化。在介电电泳分离期间，保持细胞活力可能是重要的，特别是当细胞在下游微流体组件处进行生物学测试时。
Electromanipulation of cells as a label free cell manipulation and characterization tool has gained particular interest recently. However, the applicability of electromanipulation, particularly dielectrophoresis (DEP), to biological cells is limited to cells suspended in buffers containing lower amounts of salts relative to the physiological buffers. One might question the use of low conductivity buffers (LCB) for DEP separation, as cells are stressed in buffers lacking physiological levels of salt. In LCB, cells leak ions and undergo volume regulation. Therefore, cells exhibit time-dependent DEP response in LCB. In this work, cellular changes in LCB are assessed by dielectric spectroscopy, cell viability assay, and gene expression of chondrocytes and Jurkats. Results indicate leakage of ions from cells, increases in cytoplasmic conductivity, membrane capacitance and conductance. Separability factor, which defines optimum conditions for DEP cell separation, for the two cell types is calculated using the cellular dielectric data. Optimum DEP separation conditions change as cellular dielectric properties evolve in LCB. Genetic analyses indicate no changes in expression of ionic channel proteins for chondrocytes suspended in LCB. Retaining cellular viability might be important during dielectrophoretic separation, especially when cells are to be biologically tested at a downstream microfluidic component.