Conditional immortalization of human atrial myocytes for the generation of in vitro models of atrial fibrillation.

Autor: Harlaar N; Department of Cardiology, Leiden University Medical Center, Leiden, the Netherlands.; Department of Cardiothoracic Surgery, Leiden University Medical Center, Leiden, the Netherlands., Dekker SO; Department of Cardiology, Leiden University Medical Center, Leiden, the Netherlands., Zhang J; Department of Cardiology, Leiden University Medical Center, Leiden, the Netherlands., Snabel RR; Department of Molecular Developmental Biology, Radboud University, Nijmegen, the Netherlands., Veldkamp MW; Department of Clinical and Experimental Cardiology, Amsterdam UMC, Amsterdam, the Netherlands., Verkerk AO; Department of Clinical and Experimental Cardiology, Amsterdam UMC, Amsterdam, the Netherlands.; Department of Medical Biology, Amsterdam UMC, Amsterdam, the Netherlands., Fabres CC; Applied Stem Cell Technologies, University of Twente, Enschede, the Netherlands., Schwach V; Applied Stem Cell Technologies, University of Twente, Enschede, the Netherlands., Lerink LJS; Department of Cardiology, Leiden University Medical Center, Leiden, the Netherlands., Rivaud MR; Department of Clinical and Experimental Cardiology, Amsterdam UMC, Amsterdam, the Netherlands., Mulder AA; Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, the Netherlands., Corver WE; Department of Pathology, Leiden University Medical Center, Leiden, the Netherlands., Goumans MJTH; Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, the Netherlands., Dobrev D; Institute of Pharmacology, University Duisburg-Essen, Essen, Germany., Klautz RJM; Department of Cardiothoracic Surgery, Leiden University Medical Center, Leiden, the Netherlands., Schalij MJ; Department of Cardiology, Leiden University Medical Center, Leiden, the Netherlands., Veenstra GJC; Department of Molecular Developmental Biology, Radboud University, Nijmegen, the Netherlands., Passier R; Applied Stem Cell Technologies, University of Twente, Enschede, the Netherlands., van Brakel TJ; Department of Cardiothoracic Surgery, Leiden University Medical Center, Leiden, the Netherlands., Pijnappels DA; Department of Cardiology, Leiden University Medical Center, Leiden, the Netherlands., de Vries AAF; Department of Cardiology, Leiden University Medical Center, Leiden, the Netherlands. a.a.f.de_vries@lumc.nl.
Jazyk: angličtina
Zdroj: Nature biomedical engineering [Nat Biomed Eng] 2022 Apr; Vol. 6 (4), pp. 389-402. Date of Electronic Publication: 2022 Jan 06.
DOI: 10.1038/s41551-021-00827-5
Abstrakt: The lack of a scalable and robust source of well-differentiated human atrial myocytes constrains the development of in vitro models of atrial fibrillation (AF). Here we show that fully functional atrial myocytes can be generated and expanded one-quadrillion-fold via a conditional cell-immortalization method relying on lentiviral vectors and the doxycycline-controlled expression of a recombinant viral oncogene in human foetal atrial myocytes, and that the immortalized cells can be used to generate in vitro models of AF. The method generated 15 monoclonal cell lines with molecular, cellular and electrophysiological properties resembling those of primary atrial myocytes. Multicellular in vitro models of AF generated using the immortalized atrial myocytes displayed fibrillatory activity (with activation frequencies of 6-8 Hz, consistent with the clinical manifestation of AF), which could be terminated by the administration of clinically approved antiarrhythmic drugs. The conditional cell-immortalization method could be used to generate functional cell lines from other human parenchymal cells, for the development of in vitro models of human disease.
(© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)
Databáze: MEDLINE
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