Establishment of an automated patch-clamp platform for electrophysiological and pharmacological evaluation of hiPSC-CMs
Publikation: Beitrag in Fachzeitschrift › Forschungsartikel › Beigetragen › Begutachtung
Beitragende
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 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.
Details
Originalsprache | Englisch |
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Aufsatznummer | 101662 |
Fachzeitschrift | Stem cell research |
Jahrgang | 41 |
Publikationsstatus | Veröffentlicht - Dez. 2019 |
Peer-Review-Status | Ja |
Externe IDs
PubMed | 31809994 |
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ORCID | /0000-0003-2514-9429/work/148606779 |
Schlagworte
ASJC Scopus Sachgebiete
Schlagwörter
- Automated patch-clamp, Ca current, Dynamic current-clamp, Human induced pluripotent stem cell-derived cardiomyocyte, Na current, Transient outward and inward rectifier K currents