大鼠 H9C2、人 AC16 和鼠 HL-1 心肌细胞系的转录组学分析和比较表征,Cardiovascular Research

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大鼠 H9C2、人 AC16 和鼠 HL-1 心肌细胞系的转录组学分析和比较表征
Cardiovascular Research ( IF 10.2 ) Pub Date : 2022-06-10 , DOI: 10.1093/cvr/cvac066.008
Z Gulyas-Onodi ₁ , T Visnovitz ₁ , A Koncz ₁ , B Varadi ₁ , B Agg ₁ , B Kiss ₁ , A Makkos ₁ , NR Nagy ₁ , VE Toth ₁ , P Leszek ₂ , A Gorbe ₁ , Z Giricz ₁ , EI Buzas ₁ , P Ferdinandy ₁ , ZV Varga ₁

Affiliation

资金致谢资金来源类型：公共拨款——欧盟资金。主要资金来源：欧盟地平线 2020 研究和创新计划匈牙利国家研究、发展和创新基金背景心脏细胞系和原代细胞培养物广泛用于体外模拟各种心血管疾病。尽管使用这些模型的出版物越来越多，但这些细胞系的局限性仍未确定。目的我们研究的目的是通过转录组学分析和形态学特征比较最常用的心脏细胞系与原代培养物和成熟心脏组织。方法 H9C2（大鼠）、AC16（人）和 HL-1（小鼠）心肌细胞系分化为更类似于心肌细胞的表型，通过文献中最广泛使用的方法，在增殖和分化阶段收获细胞。整个左心室组织、从小鼠和大鼠中分离的新生儿原代心肌细胞或人诱导的多能干细胞衍生的心肌细胞 (hiPSC-CM) 用作参考。在所有细胞模型上进行心脏结构蛋白的转录组分析和免疫细胞化学检测。结果与原代细胞或 hiPSC-CM 和成人组织对照相比，细胞系中心脏标志物（例如 Tnnt2、Ryr2、Tnni3）的 RNA 表达显着降低。分化程序导致 AC16 和 H9C2 系中心脏标志物显着增加和胚胎标志物减少；然而，与相应的心肌或原代培养物相比，所有细胞系中所研究基因的整体表达模式显示出显着差异。免疫细胞化学证实细胞系中结构蛋白 α-肌动蛋白和肌钙蛋白 I 的低表达。结论心肌细胞标志物和 mRNA 谱的表达模式表明，无论使用何种分化方案，细胞系与原代细胞/心脏组织的相似性为低至中度。在选择细胞作为体外平台来模拟心肌细胞和心血管疾病时，应考虑到这些限制。结论心肌细胞标志物和 mRNA 谱的表达模式表明，无论使用何种分化方案，细胞系与原代细胞/心脏组织的相似性为低至中度。在选择细胞作为体外平台来模拟心肌细胞和心血管疾病时，应考虑到这些限制。结论心肌细胞标志物和 mRNA 谱的表达模式表明，无论使用何种分化方案，细胞系与原代细胞/心脏组织的相似性为低至中度。在选择细胞作为体外平台来模拟心肌细胞和心血管疾病时，应考虑到这些限制。

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Transcriptomic analysis and comparative characterization of rat H9C2, human AC16 and murine HL-1 cardiac cell lines

Funding Acknowledgements Type of funding sources: Public grant(s) – EU funding. Main funding source(s): European Union's Horizon 2020 research and innovation programme National Research, Development and Innovation Fund of Hungary Background Cardiac cell lines and primary cell cultures are widely used to model various cardiovascular diseases in vitro. Despite the increasing number of publications using these models, limitations of these cell lines are still undetermined. Purpose The aim of our study was to compare the most commonly used cardiac cell lines to primary cultures and to mature cardiac tissues by transcriptomic analysis and morphological characterization. Methods H9C2 (rat), AC16 (human) and HL-1 (mouse) cardiac cell lines were differentiated towards a phenotype more resembling cardiomyocytes, by methods most widely used in the literature, and cells were harvested at stages of proliferation and differentiation. Whole left ventricular tissue, neonatal primary cardiac myocytes isolated from mice and rats, or human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) were applied as references. Transcriptome analysis and immunocytochemical detection of cardiac structural proteins were performed on all cell models. Results RNA expression of cardiac markers (e.g. Tnnt2, Ryr2, Tnni3) was markedly lower in cell lines compared to primary cells or hiPSC-CM and adult tissue controls. Differentiation procedures induced a significant increase in cardiac- and decrease in embryonic markers in AC16 and H9C2 lines; however, the overall expression pattern of investigated genes in all cell lines showed significant differences in comparison to corresponding myocardium or primary cultures. Immunocytochemistry confirmed low expressions of structural protein alpha-actinin and troponin I in cell lines. Conclusion Expression patterns of cardiomyocyte markers and mRNA profile indicates low-to-moderate similarity of cell lines to primary cells/cardiac tissues regardless the differentiation protocol used. These limitations should be taken into account while choosing cells as in vitro platforms to model cardiomyocytes and cardiovascular diseases.

更新日期：2022-06-10

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