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  Action potential characterization of human induced pluripotent stem cell-derived cardiomyocytes using automated patch-clamp technology.

Scheel, O., Frech, S., Amuzescu, B., Eisfeld, J., Lin, K. H., & Knott, T. (2014). Action potential characterization of human induced pluripotent stem cell-derived cardiomyocytes using automated patch-clamp technology. Assay and Drug Development Technologies, 12(8), 457-469. doi:10.1089/adt.2014.601.

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 Urheber:
Scheel, O., Autor
Frech, S., Autor
Amuzescu, B., Autor
Eisfeld, J., Autor
Lin, K. H.1, Autor           
Knott, T., Autor
Affiliations:
1Research Group of Activity-Dependent and Developmental Plasticity at the Calyx of Held, MPI for biophysical chemistry, Max Planck Society, ou_578581              

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 Zusammenfassung: Abstract Recent progress in embryonic stem cell (ESC) and induced pluripotent stem cell (iPSC) research led to high-purity preparations of human cardiomyocytes (CMs) differentiated from these two sources-suitable for tissue regeneration, in vitro models of disease, and cardiac safety pharmacology screening. We performed a detailed characterization of the effects of nifedipine, cisapride, and tetrodotoxin (TTX) on Cor.4U(®) human iPSC-CM, using automated whole-cell patch-clamp recordings with the CytoPatch(™) 2 equipment, within a complex assay combining multiple voltage-clamp and current-clamp protocols in a well-defined sequence, and quantitative analysis of several action potential (AP) parameters. We retrieved three electrical phenotypes based on AP shape: ventricular, atrial/nodal, and S-type (with ventricular-like depolarization and lack of plateau). To suppress spontaneous firing, present in many cells, we injected continuously faint hyperpolarizing currents of -10 or -20 pA. We defined quality criteria (both seal and membrane resistance over 1 GΩ), and focused our study on cells with ventricular-like AP. Nifedipine induced marked decreases in AP duration (APD): APD90 (49.8% and 40.8% of control values at 1 and 10 μM, respectively), APD50 (16.1% and 12%); cisapride 0.1 μM increased APD90 to 176.2%; and tetrodotoxin 10 μM decreased maximum slope of phase to 33.3% of control, peak depolarization potential to 76.3% of control, and shortened APD90 on average to 80.4%. These results prove feasibility of automated voltage- and current-clamp recordings on human iPSC-CM and their potential use for in-depth drug evaluation and proarrhythmic liability assessment, as well as for diagnosis and pharmacology tests for cardiac channelopathy patients.

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Sprache(n): eng - English
 Datum: 2014-10-292014-11-10
 Publikationsstatus: Erschienen
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 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.1089/adt.2014.601
 Art des Abschluß: -

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Titel: Assay and Drug Development Technologies
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: -
Seiten: - Band / Heft: 12 (8) Artikelnummer: - Start- / Endseite: 457 - 469 Identifikator: -