Institute of Human Genetics, Newcastle University, Newcastle, NE1 3BZ, UK.
In Vitro Cell Dev Biol Anim. 2011 Feb;47(2):125-31. doi: 10.1007/s11626-010-9368-1. Epub 2010 Nov 17.
Cardiac hypertrophy is a major risk factor for heart failure and associated patient morbidity and mortality. Research investigating the aberrant molecular processes that occur during cardiac hypertrophy uses primary cardiomyocytes from neonatal rat hearts as the standard experimental in vitro system. In addition, some studies make use of the H9C2 rat cardiomyoblast cell line, which has the advantage of being an animal-free alternative; however, the extent to which H9C2 cells can accurately mimic the hypertrophic responses of primary cardiac myocytes has not yet been fully established. To address this limitation, we have directly compared the hypertrophic responses of H9C2 cells with those of primary rat neonatal cardiomyocytes following stimulation with hypertrophic factors. Primary rat neonatal cardiomyocytes and H9C2 cells were cultured in vitro and treated with angiotensin II and endothelin-1 to promote hypertrophic responses. An increase in cellular footprint combined with rearrangement of cytoskeleton and induction of foetal heart genes were directly compared in both cell types using microscopy and real-time rtPCR. H9C2 cells showed almost identical hypertrophic responses to those observed in primary cardiomyocytes. This finding validates the importance of H9C2 cells as a model for in vitro studies of cardiac hypertrophy and supports current work with human cardiomyocyte cell lines for prospective molecular studies in heart development and disease.
心肌肥厚是心力衰竭以及相关患者发病率和死亡率的主要危险因素。研究心脏肥厚过程中发生的异常分子过程时,通常使用新生大鼠心脏的原代心肌细胞作为标准的体外实验系统。此外,一些研究利用 H9C2 大鼠心肌细胞系,该细胞系具有无动物替代物的优势;然而,H9C2 细胞在多大程度上能够准确模拟原代心肌细胞的肥厚反应尚未完全确定。为了解决这一限制,我们直接比较了 H9C2 细胞与原代新生大鼠心肌细胞在受到肥厚因子刺激后的肥厚反应。原代新生大鼠心肌细胞和 H9C2 细胞在体外培养,并分别用血管紧张素 II 和内皮素-1 处理以促进肥厚反应。使用显微镜和实时 rtPCR,直接比较了两种细胞类型中细胞足迹的增加,以及细胞骨架的重排和胎儿心脏基因的诱导。H9C2 细胞显示出与原代心肌细胞观察到的几乎相同的肥厚反应。这一发现验证了 H9C2 细胞作为体外心肌肥厚研究模型的重要性,并支持当前利用人类心肌细胞系进行心脏发育和疾病的潜在分子研究的工作。