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首页> 外文期刊>Review of Scientific Instruments >Novel in situ normal streaming potential device for characterizing electrostatic properties of confluent cells
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Novel in situ normal streaming potential device for characterizing electrostatic properties of confluent cells

机译：用于表征融合细胞静电特性的新型原位常规流式电势装置

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The characteristics of transport across confluent cell monolayers may often be attributed to its electrostatic properties. While tangential streaming potential is often used to quantify these electrostatic properties, this method is not effective for transport normal to the apical cell surface where the charge properties along the basolateral sides may be important (i.e., confluent cells with leaky tight junctions). In addition, even when cells have a uniform charge distribution, the shear stress generated by the conventional tangential flow device may dislodge cells from their confluent state. Here we introduce a novel streaming potential measurement device to characterize the normal electrostatic properties of confluent cells. The streaming potential device encompasses a 24 mm cell-seeded Transwell® with two AgCl electrodes on either side of the cell-seeded Transwell. Phosphate buffered saline is pressurized transversal to the Transwell and the resultant pressure gradient induces a potential difference. Confluent monolayers of HEK and EA926 cells are used as examples. The corresponding zeta potential of the cell-membrane configuration is calculated using the Helmholtz–Smoluchowski equation and the zeta potential of the confluent cell layer is deconvolved from the overall measurements. For these test models, the zeta potential is consistent with that determined using a commercial dispersed-cell device. This novel streaming potential device provides a simple, easy, and cost-effective methodology to determine the normal zeta potential of confluent cells cultured on Transwell systems while keeping the cells intact. Furthermore, its versatility allows periodic measurements of properties of the same cell culture during transient studies.
机译：跨汇合的细胞单层转运的特性通常可归因于其静电特性。尽管通常使用切向流电势来量化这些静电特性，但这种方法对于将法向垂直传输到顶端细胞表面（其中沿基底外侧的电荷特性可能很重要）（即具有泄漏紧密连接的融合细胞）无效。另外，即使当细胞具有均匀的电荷分布时，由常规切向流动装置产生的剪切应力也可能使细胞脱离其汇合状态。在这里，我们介绍了一种新颖的流电势测量设备，以表征融合细胞的正常静电性能。流电势装置包含一个24 mm的细胞播种Transwell ®，在细胞播种的Transwell的任一侧均具有两个AgCl电极。磷酸盐缓冲盐水横向加压至Transwell，由此产生的压力梯度会引起电势差。 HEK和EA926细胞的融合单层用作示例。使用Helmholtz–Smoluchowski方程计算细胞膜构型的相应zeta电位，并从总体测量值中解出融合细胞层的zeta电位。对于这些测试模型，ζ电位与使用商业分散电池设备确定的电位一致。这种新颖的流式电势设备提供了一种简单，简便且具有成本效益的方法，可确定在Transwell系统上培养的融合细胞的正常Zeta电势，同时保持细胞完整。此外，它的多功能性允许在瞬态研究期间定期测量同一细胞培养物的特性。

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