Abstract：
BACKGROUND: The conventional \"whole-cell patch-clamp\" recording technique is widely used to measure the resting membrane potential (VM) and to dissect the underlying membrane ionic conductances in isolated vascular endothelial cells.
METHODS: Herein, we assessed whether the automated patch-clamp (APC) technology, which replaces the traditional patch-pipette with a planar substrate to permit researchers lacking formal training in electrophysiology to generate large amounts of data in a relatively short time, can be used to characterize the bioelectrical activity of vascular endothelial cells. We assessed whether the Port-a-Patch planar patch-clamp system, which is regarded as the smallest electrophysiological rig available on the market, can be used to measure the VM and resting membrane currents in the human cerebrovascular endothelial cell line, hCMEC/D3.
METHODS: We demonstrated that the Port-a-Patch planar patch-clamp system provides the same values of the resting VM as those provided by the conventional patch-clamp technique. Furthermore, the APC technology provides preliminary data demonstrating that the resting VM of hCMEC/D3 cells is primarily contributed by Cl- and Na+, as demonstrated with the patch-clamp technique for many other endothelial cell types.
CONCLUSIONS: The Port-a-Patch planar patch-clamp system can be successfully used to measure the resting VM and the underlying membrane ionic conductances in hCMEC/D3 cells. We envisage that this easy-to-use APC system could also be extremely useful for the investigation of the membrane currents that can be activated by chemical, thermal, optical, and mechanical stimuli in this cell line as well as in other types of isolated vascular endothelial cells.
METHODS: Herein, we assessed whether the automated patch-clamp (APC) technology, which replaces the traditional patch-pipette with a planar substrate to permit researchers lacking formal training in electrophysiology to generate large amounts of data in a relatively short time, can be used to characterize the bioelectrical activity of vascular endothelial cells. We assessed whether the Port-a-Patch planar patch-clamp system, which is regarded as the smallest electrophysiological rig available on the market, can be used to measure the VM and resting membrane currents in the human cerebrovascular endothelial cell line, hCMEC/D3.
METHODS: We demonstrated that the Port-a-Patch planar patch-clamp system provides the same values of the resting VM as those provided by the conventional patch-clamp technique. Furthermore, the APC technology provides preliminary data demonstrating that the resting VM of hCMEC/D3 cells is primarily contributed by Cl- and Na+, as demonstrated with the patch-clamp technique for many other endothelial cell types.
CONCLUSIONS: The Port-a-Patch planar patch-clamp system can be successfully used to measure the resting VM and the underlying membrane ionic conductances in hCMEC/D3 cells. We envisage that this easy-to-use APC system could also be extremely useful for the investigation of the membrane currents that can be activated by chemical, thermal, optical, and mechanical stimuli in this cell line as well as in other types of isolated vascular endothelial cells.
摘要：
背景：传统的“全细胞膜片钳”记录技术被广泛用于测量静息膜电位（VM）和解剖离体血管内皮细胞中潜在的膜离子电导。
方法：这里，我们评估了自动膜片钳（APC）技术，用平面基板代替传统的贴片移液器，以允许缺乏电生理学正式培训的研究人员在相对较短的时间内生成大量数据，可用于表征血管内皮细胞的生物电活性。我们评估了Port-a-Patch平面膜片钳系统是否，被认为是市场上最小的电生理钻机，可用于测量人脑血管内皮细胞系中的VM和静息膜电流，hCMEC/D3。
方法：我们证明了Port-a-Patch平面膜片钳系统提供的静息VM值与常规膜片钳技术提供的值相同。此外，APC技术提供的初步数据表明，hCMEC/D3细胞的静息VM主要由Cl-和Na+贡献，正如许多其他类型的内皮细胞的膜片钳技术所证明的那样。
结论：Port-a-Patch平面膜片钳系统可成功用于测量hCMEC/D3细胞中的静息VM和基础膜离子电导。我们设想这种易于使用的APC系统对于研究可以通过化学激活的膜电流也非常有用，热,以及该细胞系以及其他类型的分离血管内皮细胞中的机械刺激。
方法：这里，我们评估了自动膜片钳（APC）技术，用平面基板代替传统的贴片移液器，以允许缺乏电生理学正式培训的研究人员在相对较短的时间内生成大量数据，可用于表征血管内皮细胞的生物电活性。我们评估了Port-a-Patch平面膜片钳系统是否，被认为是市场上最小的电生理钻机，可用于测量人脑血管内皮细胞系中的VM和静息膜电流，hCMEC/D3。
方法：我们证明了Port-a-Patch平面膜片钳系统提供的静息VM值与常规膜片钳技术提供的值相同。此外，APC技术提供的初步数据表明，hCMEC/D3细胞的静息VM主要由Cl-和Na+贡献，正如许多其他类型的内皮细胞的膜片钳技术所证明的那样。
结论：Port-a-Patch平面膜片钳系统可成功用于测量hCMEC/D3细胞中的静息VM和基础膜离子电导。我们设想这种易于使用的APC系统对于研究可以通过化学激活的膜电流也非常有用，热,以及该细胞系以及其他类型的分离血管内皮细胞中的机械刺激。
