scholarly journals Action Potential Characterization of Human Induced Pluripotent Stem Cell–Derived Cardiomyocytes Using Automated Patch-Clamp Technology

2014 ◽  
Vol 12 (8) ◽  
pp. 457-469 ◽  
Author(s):  
Olaf Scheel ◽  
Stefanie Frech ◽  
Bogdan Amuzescu ◽  
Jörg Eisfeld ◽  
Kun-Han Lin ◽  
...  
Keyword(s):  
Stem Cell ◽  
Patch Clamp ◽  
Action Potential ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Automated Patch Clamp ◽  
Induced Pluripotent

scholarly journals Characterizing Human Ion Channels in Induced Pluripotent Stem Cell–Derived Neurons

2012 ◽  
Vol 17 (9) ◽  
pp. 1264-1272 ◽  
Author(s):  
Alison Haythornthwaite ◽  
Sonja Stoelzle ◽  
Alexander Hasler ◽  
Andrea Kiss ◽  
Johannes Mosbacher ◽  
...  
Keyword(s):  
Stem Cell ◽  
Ion Channels ◽  
Patch Clamp ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Excitable Cells ◽  
Automated Patch Clamp ◽  
Induced Pluripotent ◽  
Two Populations ◽  
A Current

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.

scholarly journals Automated Patch-Clamp Pharmacology Assays using Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

2015 ◽  
Vol 108 (2) ◽  
pp. 585a
Author(s):  
Olaf Scheel ◽  
Stefanie Frech ◽  
Bogdan P. Amuzescu ◽  
Jörg Eisfeld ◽  
Kun-Han Lin ◽  
...  
Keyword(s):  
Stem Cell ◽  
Patch Clamp ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Automated Patch Clamp ◽  
Induced Pluripotent

scholarly journals A Temperature-Controlled Patch Clamp Platform Demonstrated on Jurkat T Lymphocytes and Human Induced Pluripotent Stem Cell-Derived Neurons

2020 ◽  
Vol 7 (2) ◽  
pp. 46 ◽  
Author(s):  
Jann Harberts ◽  
Max Kusch ◽  
John O’Sullivan ◽  
Robert Zierold ◽  
Robert H. Blick
Keyword(s):  
Stem Cell ◽  
T Lymphocytes ◽  
Patch Clamp ◽  
Pluripotent Stem Cell ◽  
Room Temperature ◽  
Standard Procedure ◽  
Induced Pluripotent Stem Cell ◽  
Electrophysiological Properties ◽  
Temperature Controlled ◽  
Induced Pluripotent

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.

Sign in / Sign up

Export Citation Format

Share Document