@article{d8432261b6d74ca4987a541e3dcb6c5e,
title = "FUCCI-Based Live Imaging Platform Reveals Cell Cycle Dynamics and Identifies Pro-proliferative Compounds in Human iPSC-Derived Cardiomyocytes",
abstract = "One of the major goals in cardiac regeneration research is to replace lost ventricular tissue with new cardiomyocytes. However, cardiomyocyte proliferation drops to low levels in neonatal hearts and is no longer efficient in compensating for the loss of functional myocardium in heart disease. We generated a human induced pluripotent stem cell (iPSC)-derived cardiomyocyte-specific cell cycle indicator system (TNNT2-FUCCI) to characterize regular and aberrant cardiomyocyte cycle dynamics. We visualized cell cycle progression in TNNT2-FUCCI and found G2 cycle arrest in endoreplicating cardiomyocytes. Moreover, we devised a live-cell compound screening platform to identify pro-proliferative drug candidates. We found that the alpha-adrenergic receptor agonist clonidine induced cardiomyocyte proliferation in vitro and increased cardiomyocyte cell cycle entry in neonatal mice. In conclusion, the TNNT2-FUCCI system is a versatile tool to characterize cardiomyocyte cell cycle dynamics and identify pro-proliferative candidates with regenerative potential in the mammalian heart.",
keywords = "cardiomyocyte, cell cycle activity, cell cycle indicator, clonidine, fluorescence ubiquitination cell cycle indicator, induced pluripotent stem cell, polyploidy",
author = "Francesca Murganti and Wouter Derks and Marion Baniol and Irina Simonova and Palina Trus and Katrin Neumann and Shahryar Khattak and Kaomei Guan and Olaf Bergmann",
note = "Funding Information: OB was supported by the Center for Regenerative Therapies Dresden, the Karolinska Institute, the Swedish Research Council, the Ragnar S{\"o}derberg Foundation, the {\AA}ke Wiberg Foundation, and the LeDucq Foundation. Funding Information: The CRTD5-TNNT2-FUCCI reporter hiPSC line was generated by the Stem Cell Engineering Facility, a core facility of the Center for Molecular and Cellular Bioengineering (CMCB) at Technische Universit{\"a}t Dresden. Giemsa staining and analysis of metaphase chromosomes were performed at the Molecular Cytogenetics lab at Jena University Hospital. We thank Marc Bickle from MPI-CBG in Dresden for advice on image acquisition strategies and for helping to set up the analysis in KNIME. We thank the Light Microscopy and Flow Cytometry core facilities of BIOTEC/CRTD for their help with imaging and flow cytometry analyses. A preprint of this article was published at bioRxiv: (57). Publisher Copyright: Copyright {\textcopyright} 2022 Murganti, Derks, Baniol, Simonova, Trus, Neumann, Khattak, Guan and Bergmann.",
year = "2022",
month = apr,
day = "25",
doi = "10.3389/fcvm.2022.840147",
language = "English",
volume = "9",
journal = "Frontiers in Cardiovascular Medicine",
issn = "2297-055X",
publisher = "Frontiers Media S.A.",
}