Zsófia Onódi1, Tamás Visnovitz2, Bernadett Kiss3, Szabolcs Hambalkó4, Anna Koncz2, Bence Ágg5, Barnabás Váradi4, Viktória É Tóth1, Regina N Nagy3, Tamás G Gergely1, Dorottya Gergő6, András Makkos3, Csilla Pelyhe4, Nóra Varga7, Dóra Reé7, Ágota Apáti7, Przemyslaw Leszek8, Tamás Kovács9, Nándor Nagy9, Péter Ferdinandy5, Edit I Buzás10, Anikó Görbe5, Zoltán Giricz5, Zoltán V Varga11. 1. Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary; HCEMM-SU Cardiometabolic Immunology Research Group, Budapest, Hungary; MTA-SE Momentum Cardio-Oncology and Cardioimmunology Research Group, Budapest, Hungary. 2. Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary. 3. Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary; MTA-SE System Pharmacology Research Group, Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary. 4. Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary. 5. Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary; MTA-SE System Pharmacology Research Group, Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary; Pharmahungary Group, Szeged, Hungary. 6. Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary; HCEMM-SU Cardiometabolic Immunology Research Group, Budapest, Hungary. 7. Research Centre for Natural Sciences, Institute of Enzymology, Budapest, Hungary; ELKH-Research Centre for Natural Sciences, Institute of Enzymology, Budapest, Hungary. 8. Department of Heart Failure and Transplantology, Cardinal Stefan Wyszyński National Institute of Cardiology, Warszawa, Poland. 9. Department of Anatomy, Histology and Embryology, Semmelweis University, Budapest, Hungary. 10. Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary; HCEMM-SU Extracellular Vesicle Research Group, Hungary; ELKH-SE Immune-Proteogenomics Extracellular Vesicle Research Group, Hungary. 11. Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest, Hungary; HCEMM-SU Cardiometabolic Immunology Research Group, Budapest, Hungary; MTA-SE Momentum Cardio-Oncology and Cardioimmunology Research Group, Budapest, Hungary. Electronic address: varga.zoltan@med.semmelweis-univ.hu.
Abstract
BACKGROUND: Cardiac cell lines and primary cells are widely used in cardiovascular research. Despite increasing number of publications using these models, comparative characterization of these cell lines has not been performed, therefore, their limitations are undetermined. We aimed to compare cardiac cell lines to primary cardiomyocytes and to mature cardiac tissues in a systematic manner. METHODS AND RESULTS: Cardiac cell lines (H9C2, AC16, HL-1) were differentiated with widely used protocols. Left ventricular tissue, neonatal primary cardiomyocytes, and human induced pluripotent stem cell-derived cardiomyocytes served as reference tissue or cells. RNA expression of cardiac markers (e.g. Tnnt2, Ryr2) was markedly lower in cell lines compared to references. Differentiation induced increase in cardiac- and decrease in embryonic markers however, the overall transcriptomic profile and annotation to relevant biological processes showed consistently less pronounced cardiac phenotype in all cell lines in comparison to the corresponding references. Immunocytochemistry confirmed low expressions of structural protein sarcomeric alpha-actinin, troponin I and caveolin-3 in cell lines. Susceptibility of cell lines to sI/R injury in terms of viability as well as mitochondrial polarization differed from the primary cells irrespective of their degree of differentiation. CONCLUSION: Expression patterns of cardiomyocyte markers and whole transcriptomic profile, as well as response to sI/R, and to hypertrophic stimuli indicate low-to-moderate similarity of cell lines to primary cells/cardiac tissues regardless their differentiation. Low resemblance of cell lines to mature adult cardiac tissue limits their potential use. Low translational value should be taken into account while choosing a particular cell line to model cardiomyocytes.
BACKGROUND: Cardiac cell lines and primary cells are widely used in cardiovascular research. Despite increasing number of publications using these models, comparative characterization of these cell lines has not been performed, therefore, their limitations are undetermined. We aimed to compare cardiac cell lines to primary cardiomyocytes and to mature cardiac tissues in a systematic manner. METHODS AND RESULTS: Cardiac cell lines (H9C2, AC16, HL-1) were differentiated with widely used protocols. Left ventricular tissue, neonatal primary cardiomyocytes, and human induced pluripotent stem cell-derived cardiomyocytes served as reference tissue or cells. RNA expression of cardiac markers (e.g. Tnnt2, Ryr2) was markedly lower in cell lines compared to references. Differentiation induced increase in cardiac- and decrease in embryonic markers however, the overall transcriptomic profile and annotation to relevant biological processes showed consistently less pronounced cardiac phenotype in all cell lines in comparison to the corresponding references. Immunocytochemistry confirmed low expressions of structural protein sarcomeric alpha-actinin, troponin I and caveolin-3 in cell lines. Susceptibility of cell lines to sI/R injury in terms of viability as well as mitochondrial polarization differed from the primary cells irrespective of their degree of differentiation. CONCLUSION: Expression patterns of cardiomyocyte markers and whole transcriptomic profile, as well as response to sI/R, and to hypertrophic stimuli indicate low-to-moderate similarity of cell lines to primary cells/cardiac tissues regardless their differentiation. Low resemblance of cell lines to mature adult cardiac tissue limits their potential use. Low translational value should be taken into account while choosing a particular cell line to model cardiomyocytes.
Authors: Max J Cumberland; Leto L Riebel; Ashwin Roy; Christopher O'Shea; Andrew P Holmes; Chris Denning; Paulus Kirchhof; Blanca Rodriguez; Katja Gehmlich Journal: Front Physiol Date: 2022-02-07 Impact factor: 4.566
Authors: Ivana Fiserova; Minh Duc Trinh; Moustafa Elkalaf; Lukas Vacek; Marek Heide; Stanislava Martinkova; Kamila Bechynska; Vit Kosek; Jana Hajslova; Ondrej Fiser; Petr Tousek; Jan Polak Journal: Front Cardiovasc Med Date: 2022-08-22