Three-Dimensional Culture Alters Primary Cardiac Cell Phenotype
Three-Dimensional Culture Alters Primary Cardiac Cell Phenotype
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DOI：
10.1089/ten.tea.2009.0458
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发表时间：
2010-02-01
影响因子：
4.1
通讯作者：
Pizarro, Christian
中科院分区：
文献类型：
--
作者：
Akins, Robert E., Jr.;Rockwood, Danielle;Pizarro, Christian
定向形成复杂的三维组织结构是组织工程和再生医学的基本目标。细胞在3D结构中的生长被认为会影响细胞的表型和功能，特别是相对细胞的分布、表达谱和对外界信号的反应；然而，相对较少的研究考察3D重组对关键靶器官细胞的影响，如心脏。因此，我们使用无血清培养液在3D模型系统中培养原代心肌细胞，以测试在3D与2D培养中生长的稳定细胞群体的表达谱、多细胞组织途径、组织成熟标志物和对激素刺激的反应性显著改变的假设。我们发现，与包含相同细胞群体的平行2D培养相比，3D培养中形成了明显的多细胞结构，伴随着mRNA表达谱的变化，内皮细胞迁移路径的上调，胎儿基因(NPPA和Ankrd1)的表达减少，以及对三碘甲腺原氨酸刺激的敏感性增加。这些结果表明，原代心肌细胞在3D聚集体中的培养导致成分细胞表型发生生理学上的变化，这与心室组织的形成和成熟相一致。
The directed formation of complex three-dimensional (3D) tissue architecture is a fundamental goal in tissue engineering and regenerative medicine. The growth of cells in 3D structures is expected to influence cellular phenotype and function, especially relative cell distribution, expression profiles, and responsiveness to exogenous signals; however, relatively few studies have been carried out to examine the effects of 3D reaggregation on cells from critical target organs, like the heart. Accordingly, we cultured primary cardiac ventricular cells in a 3D model system using a serum-free medium to test the hypothesis that expression profiles, multicellular organizational pathways, tissue maturation markers, and responsiveness to hormone stimulation were significantly altered in stable cell populations grown in 3D versus 2D culture. We found that distinct multi-cellular structures formed in 3D in conjunction with changes in mRNA expression profile, up-regulation of endothelial cell migratory pathways, decreases in the expression of fetal genes (Nppa and Ankrd1), and increased sensitivity to tri-iodothyronine stimulation when compared to parallel 2D cultures comprising the same cell populations. These results indicate that the culture of primary cardiac cells in 3D aggregates leads to physiologically relevant alterations in component cell phenotype consistent with cardiac ventricular tissue formation and maturation.