Objective Due to high-throughput capabilities, automated patch clamp system has been widely used in cardiovascular ion channel research in rescent years. The aim of this study is to establish the method for high-throughput K⁺ outward current recording on jurkat cell using automated patch clamp system.

Methods Using NPC-16 chip by which 8 cells could be recorded simultaneously, whole cell K⁺ outward current on Jurkat cell was recorded. Besides, inhibitory effects of Tetraethylammonium chloride (TEA) were monitored and IC₅₀ for TEA was also determined. Optimization of cell culture condition and seal enhancer solution containing different Ca²⁺ concentrations (10, 20, 30, 35, 40, 45 mM respectively) was performed to improve success rate of sealing and whole-cell access.

Results For 96 cells from NPC-16 chips, the success rate of giga-seal and whole-cell access was 95% and 87% respectively. For cell culture optimization, efficient cell passaging by dissociating cell clusters to single cells could facilitate seal and whole-cell access. Among different concentration of Ca²⁺, 30 mM Ca²⁺ was found to be most efficient for giga-seal and whole-cell access. Moreover, The K⁺ outward current could be inhibited by TEA in a dose dependent manner with IC₅₀ (10.6 ± 2.7 mM).

Conclusions Taken together, one method for high-throughput K⁺ outward current recording on Jurkat cell by automated patch clamp system was successfully established with high success rate of giga-seals and whole-cell access. Optimization of cell culture condition and seal enhancer solution could promote higher success rate. The system will allows for a cost-effective and high throughout cell currents recording in future cardiovascular disease research.