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2019 - Establishment of an automated patch-clamp platform for electrophysiological and pharmacological evaluation of hiPSC-CMs

icon pl   Patchliner and Dynamite8 publication in Stem Cell Research (2019)

Authors:
Li W., Luo X., Y., Wagner M., Piorkowski C., El-Armouche A., Guan K.

Journal:
Stem Cell Research (2019) In Press, Journal Pre-proof doi.org/10.1016/j.scr.2019.101662


Abstract:

Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) have evolved into widely used and reliable cell sources for modeling cardiovascular channelopathies and for drug safety pharmacology. However, the electrophysiological and pharmacological applications of hiPSC-CMs are hampered by manual patch-clamp technique, which is labor-intensive and generates a low-output. The automated patch-clamp technique is showing potential to overcome this problem. Here, we describe a new dissociation method, with which we can harvest a vast number of single relaxed hiPSC-CMs with smooth membrane suited for automated patch-clamp. Using the automated whole-cell patch-clamp technology, we report a high success rate for cell capture and whole-cell access (around 70%). We are able to identify and record several currents and paced action potentials (APs) with different success rates, including Na+ current (INa), L-type Ca2+ current (ICaL), two specific K+ currents, the transient outward K+ current (Ito) and the inward rectifier K+ current (IK1). Moreover, we successfully applied dynamic current-clamp to virtually increase IK1 for AP recordings. Our study suggests that automated patch-clamp technology could be used to investigate the relevant ionic currents and APs in hiPSC-CMs. The combination of automated patch-clamp and hiPSC-CM technologies promises a wide range of applications in the future.

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