Systematic transcriptomic and phenotypic characterization of human and murine cardiac myocyte cell lines and primary cardiomyocytes reveals serious limitations and low resemblances to adult cardiac phenotype. (April 2022)

• Record Type: Journal Article
• Authors: Onódi, Zsófia; Visnovitz, Tamás; Kiss, Bernadett; Hambalkó, Szabolcs; Koncz, Anna; Ágg, Bence; Váradi, Barnabás; Tóth, Viktória É.; Nagy, Regina N.; Gergely, Tamás G.; Gergő, Dorottya; Makkos, András; Pelyhe, Csilla; Varga, Nóra; Reé, Dóra; Apáti, Ágota; Leszek, Przemyslaw; Kovács, Tamás; Nagy, Nándor; Ferdinandy, Péter; Buzás, Edit I.; Görbe, Anikó; Giricz, Zoltán; Varga, Zoltán V.
• Is Part Of: Journal of molecular and cellular cardiology. Volume 165(2022)
• Page Start: 19
• Page End: 30
• Publication Date: 2022-04
• Subjects: H9C2 -- AC16 -- HL-1 -- Cardiomyocyte -- Stem cell -- Primary cell culture; hiPSC-CM human induced pluripotent stem cell-derived cardiac myocyte -- NRCM neonatal rat cardiac myocyte -- NMCM neonatal murine cardiac myocyte -- AC16/H9C2 Diff differentiated from of AC16/H9C2 -- ATRA all-trans retinoic acid -- DEG differentially expressed genes -- PCA principal component analysis -- GO gene ontology -- sI/R simulated ischemia-reperfusion -- SV40 simian virus 40 -- ANGII angiotensin II -- ISOP isoprenaline -- ITS insulin-transferrin selenium supplement -- FBS fetal bovine serum; Cardiology -- Periodicals; Heart Diseases -- Periodicals; Molecular Biology -- Periodicals; Cardiologie -- Périodiques; Cardiology; Electronic journals; Periodicals; 616.12
• DOI: 10.1016/j.yjmcc.2021.12.007
• Languages: English
• ISSNs: 0022-2828

Abstract

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. Graphical abstract: Unlabelled Image Highlights: Immortalized cardiac cell lines show low resemblance to mature cardiac cells. Organized sarcomeric structure is detectable in HL-1 cells but not in H9C2 or AC16. Differentiation of AC16 or H9C2 induces minor changes towards cardiac phenotype. Functional tests reveal the restricted responsiveness of all the cardiac cell lines. This is the first systematic study showing major limitations of cardiac cell lines.