TY - JOUR
T1 - Conditional immortalization of human atrial myocytes for the generation of in vitro models of atrial fibrillation
AU - Harlaar, Niels
AU - Dekker, Sven O.
AU - Zhang, Juan
AU - Snabel, Rebecca R.
AU - Veldkamp, Marieke W.
AU - Verkerk, Arie O.
AU - Fabres, Carla Cofiño
AU - Schwach, Verena
AU - Lerink, Lente J.S.
AU - Rivaud, Mathilde R.
AU - Mulder, Aat A.
AU - Corver, Willem E.
AU - Goumans, Marie José T.H.
AU - Dobrev, Dobromir
AU - Klautz, Robert J.M.
AU - Schalij, Martin J.
AU - Veenstra, Gert Jan C.
AU - Passier, Robert
AU - van Brakel, Thomas J.
AU - Pijnappels, Daniël A.
AU - de Vries, Antoine A.F.
N1 - Funding Information: We thank J. Liu (LUMC, Leiden, the Netherlands) for constructing plasmid pLV. iMHCK7.SV40-LT-WT and producing LV.iMHCK7.SV40-LT-WT particles, T. v. Herwaarden (LUMC, Leiden, the Netherlands) for collecting human foetal atrial tissue, B. Klein (LUMC, Leiden, the Netherlands) for donating the SV40 LT-encoding plasmid pAT153.SV40ori, S. Hauschka (University of Washington, Seattle, WA) for providing the construct +aMHCKChCAT encoding the MHCK7 promoter, D. Trono (Swiss Federal Institute of Technology Lausanne, Lausanne, Switzerland) for making available the LV shuttle plasmid pLVET-tTR-KRAB, C. Jost (LUMC, Leiden, the Netherlands) for assistance with interpretation and annotation of transmission electron microscopy data and U. Ravens (University of Freiburg, Freiburg, Germany) for useful discussions.This publication received financial support from the LUMC Executive Board (PhD fellowship to N.H.) and from the Leiden Regenerative Medicine Platform Holding (LRMPH project 8212/41235 to A.A.F.d.V.). Additional support was provided by the research programme ‘More Knowledge with Fewer Animals’ (MKMD, project 114022503, to A.A.F.d.V.), which is (partly) financed by the Netherlands Organisation for Health Research and Development (ZonMw) and by the Dutch Society for the Replacement of Animal Testing (dsRAT). Publisher Copyright: © 2022, The Author(s), under exclusive licence to Springer Nature Limited.
PY - 2022/4
Y1 - 2022/4
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85122411856&partnerID=8YFLogxK
U2 - https://doi.org/10.1038/s41551-021-00827-5
DO - https://doi.org/10.1038/s41551-021-00827-5
M3 - Article
C2 - 34992271
SN - 2157-846X
VL - 6
SP - 389
EP - 402
JO - Nature Biomedical Engineering
JF - Nature Biomedical Engineering
IS - 4
ER -