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2019 - Introducing simulated IK1 into human iPSC-cardiomyocytes using dynamic clamp on an automated patch clamp setup

 icon pl   Patchliner (dynamic clamp) poster, BPS Meeting 2019  logo pdf   (1.3 MB)

 

Abstract:

Dynamic clamp is a powerful tool to inject real-time simulated currents into patch clamped cells1. This has been shown using conventional patch clamp whereby the inward rectifier current IK1 was introduced into human stem cell-derived cardiomyocytes (hSC-CMs)2,3. IK1 is expressed at low levels in these cells, hence their membrane potential is more depolarized than that of primary cardiomyocytes, limiting their use in safety pharmacology. Introducing simulated IK1 into hSC-CMs may render them a viable alternative to scarcely available adult human cardiomyocytes4,5. In this study, we combined dynamic clamp with an automated patch clamp (APC) system to demonstrate that IK1 conductance can be added to hSC-CMs on this platform, while at the same time, applying automatic Rseal compensation (SC). Our results show that virtual IK1 can be successfully injected into hSC-CMs in up to 8 cells simultaneously and that Rseal is correctly compensated avoiding overcompensation. Our approach results in more stable resting membrane potentials and improved action potential (AP) shape. „L”-Type calcium channel opener BayK and channel blocker nifedipine were also tested..

 

References:

1. Wilders R. (2006) J. Physiol. 576:349–359 

2. Bett GC, et al. (2013) Heart Rhythm. 10:1903–1910. 

3. Meijer van Putten RM, et al. (2015) Front. Physiol. 6:7. doi: 10.3389/fphys.2015.00007;

4. Jonsson MKB, et al. (2012) J. Mol. Cell Cardiol. 52:998–1008

5. Rajamohan D, et al. (2016) Stem Cells Dev. 25:439–452

 

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