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.
Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary.
J Mol Cell Cardiol. 2022 Apr;165:19-30. doi: 10.1016/j.yjmcc.2021.12.007. Epub 2021 Dec 24.
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.
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.
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.
心脏细胞系和原代细胞广泛应用于心血管研究。尽管使用这些模型的出版物越来越多，但尚未对这些细胞系进行比较特征描述，因此其局限性尚不确定。我们旨在以系统的方式比较心脏细胞系与原代心肌细胞和成熟的心脏组织。
使用广泛使用的方案对心脏细胞系（H9C2、AC16、HL-1）进行分化。左心室组织、新生儿原代心肌细胞和人诱导多能干细胞衍生的心肌细胞作为参考组织或细胞。与参考相比，细胞系中的心脏标志物（例如 Tnnt2、Ryr2）的 RNA 表达明显较低。分化诱导增加了心脏标志物，减少了胚胎标志物，但整体转录组谱和注释到相关生物学过程表明，与相应的参考相比，所有细胞系的心脏表型始终不那么明显。免疫细胞化学证实细胞系中结构蛋白肌球蛋白重链、肌钙蛋白 I 和窖蛋白-3 的表达较低。细胞系对缺血再灌注损伤的敏感性，无论是在活力方面，还是在线粒体极化方面，都与原代细胞不同，而与分化程度无关。
心肌细胞标志物的表达模式和整个转录组谱，以及对缺血再灌注和肥厚刺激的反应，表明细胞系与原代细胞/心脏组织的相似性低至中度，无论其分化程度如何。细胞系与成熟的成人心脏组织的相似性低限制了其潜在用途。在选择特定的细胞系来模拟心肌细胞时，应考虑低的转化价值。
