scholarly journals Patch-Clamp Recording from Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes: Improving Action Potential Characteristics through Dynamic Clamp

2017 ◽  
Vol 18 (9) ◽  
pp. 1873 ◽  
Author(s):  
Arie Verkerk ◽  
Christiaan Veerman ◽  
Jan Zegers ◽  
Isabella Mengarelli ◽  
Connie Bezzina ◽  
...  
2020 ◽  
Vol 7 (2) ◽  
pp. 46 ◽  
Author(s):  
Jann Harberts ◽  
Max Kusch ◽  
John O’Sullivan ◽  
Robert Zierold ◽  
Robert H. Blick

Though patch clamping at room temperature is a widely disseminated standard procedure in the electrophysiological community, it does not represent the biological system in mammals at around 37 °C. In order to better mimic the natural environment in electrophysiological studies, we present a custom-built, temperature-controlled patch clamp platform for upright microscopes, which can easily be adapted to any upright patch clamp setup independently, whether commercially available or home built. Our setup can both cool and heat the platform having only small temperature variations of less than 0.5 °C. We demonstrate our setup with patch clamp measurements at 36 °C on Jurkat T lymphocytes and human induced pluripotent stem cell-derived neurons. Passive membrane parameters and characteristic electrophysiological properties, such as the gating properties of voltage-gated ion channels and the firing of action potentials, are compared to measurements at room temperature. We observe that many processes that are not explicitly considered as temperature dependent show changes with temperature. Thus, we believe in the need of a temperature control in patch clamp measurements if improved physiological conditions are required. Furthermore, we advise researchers to only compare electrophysiological results directly that have been measured at similar temperatures since small variations in cellular properties might be caused by temperature alterations.


2012 ◽  
Vol 17 (9) ◽  
pp. 1264-1272 ◽  
Author(s):  
Alison Haythornthwaite ◽  
Sonja Stoelzle ◽  
Alexander Hasler ◽  
Andrea Kiss ◽  
Johannes Mosbacher ◽  
...  

Neurons derived from human-induced pluripotent stem cells were characterized using manual and automated patch-clamp recordings. These cells expressed voltage-gated Na+ (Nav), Ca2+ (Cav), and K+ (Kv) channels as expected from excitable cells. The Nav current was TTX sensitive, IC50 = 12 ± 6 nM ( n = 5). About 50% of the Cav current was blocked by 10 µM of the L-type channel blocker nifedipine. Two populations of the Kv channel were present in different proportions: an inactivating (A-type) and a noninactivating type. The A-type current was sensitive to 4-AP and TEA (IC50 = 163 ± 93 µM; n = 3). Application of γ-aminobutyric acid (GABA) activated a current sensitive to the GABAA receptor antagonist bicuculline, IC50 = 632 ± 149 nM ( n = 5). In both devices, comparable action potentials were generated in the current clamp. With unbiased, automated patch clamp, about 40% of the cells expressed Nav currents, whereas visual guidance in manual patch clamp provided almost a 100% success rate of patching “excitable cells.” These results show high potential for pluripotent stem cell–derived neurons as a useful model for drug discovery, in combination with automated patch-clamp recordings for high-throughput and high-quality drug assessments at human neuronal ion channels in their correct cellular background.


2021 ◽  
Vol 2 (4) ◽  
pp. 100859
Author(s):  
Joe Z. Zhang ◽  
Shane Rui Zhao ◽  
Chengyi Tu ◽  
Paul Pang ◽  
Mao Zhang ◽  
...  

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