改良的适合膜片钳试验的血管内皮细胞处理方法

Modification of vascular endothelial cell treatment for patch clamp study

目的建立一种简便的、适合膜片钳试验的人脐静脉内皮细胞(ECV304)处理方法,并在培养的细胞上观察其钙激活钾通道(Kca)特性。方法采用改良的内皮细胞培养方法,用单通道记录法记录内皮细胞Kca电流。结果内皮细胞在光镜下呈梭形,并可以检测到细胞膜上的Kca电流。结论本方法改良了传统的内皮细胞培养方法,能在较短时间内完成膜片钳通道记录,是一种适合膜片钳试验的处理方法。

A modified protocol for quick and economic treatment of cultured human umbilical vein endothelial cells has been estabfishe, which result in high viability of the cells to allow better performance in patch-clamp studies. The electrophysiological properties of Ca^2＋ -activated K＋ （KCa） channel of the cultured cells were investigated with a cell-attached configuration. With this modified treatment method, the cultured cells appear fusiform in shape under microscope, and KCa channel currents could be detected readily, suggesting their eligibility for patch-clamp studies.