Aligned nanofiber scaffolds improve functionality of cardiomyocytes differentiated from human induced pluripotent stem cell-derived cardiac progenitor cells
Aligned nanofiber scaffolds improve functionality of cardiomyocytes differentiated from human induced pluripotent stem cell-derived cardiac progenitor cells
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DOI：
10.1038/s41598-020-70547-4
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发表时间：
2020-08-11
影响因子：
4.6
通讯作者：
Drowley, Lauren
中科院分区：
文献类型：
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作者：
Ding, Mei;Andersson, Henrik;Drowley, Lauren
心脏祖细胞（cardiac progenitor cells，CPC）具有分化为心肌细胞（cardiomyocytes，CMs）、内皮细胞（endothelial cells，ECCs）和平滑肌细胞（smooth muscle cells，smooth muscle cells）的能力，是心脏修复/再生的理想候选细胞。在体外模型系统中，细胞在更类似于体内的环境中生长，如3D培养，已被证明比2D培养更能预测细胞生物学和疾病病理生理学。在这份报告中，我们专注于使用Wnt抑制剂，通过测量标记蛋白和基因表达以及细胞内Ca 2+振荡来研究人iPSC-CPC在2D或3D培养条件下的分化。我们的结果显示，具有对齐的支架的3D培养物，模仿心脏的细胞外基质的结构，改善了iPSC-CPC向由Wnt抑制诱导的功能性心肌细胞的分化，如心肌肌钙蛋白T（cTnT）阳性细胞的数量增加和同步的细胞内Ca 2+振荡所示。此外，我们研究了是否可以将3D微生物培养物用作体外模型，通过测试许多其他分化因子（包括ALK 5抑制剂和BMP信号传导抑制剂）来进行化合物筛选。这项工作强调了使用更相关的体外模型的重要性，不仅测量标记蛋白的表达，而且测量筛选中的功能读数，以确定最佳化合物并研究所产生的生物学。
Cardiac progenitor cells (CPCs), capable of differentiating into multiple cardiac cell types including cardiomyocytes (CMs), endothelial cells, and smooth muscle cells, are promising candidates for cardiac repair/regeneration. In vitro model systems where cells are grown in a more in vivo-like environment, such as 3D cultures, have been shown to be more predictive than 2D culture for studying cell biology and disease pathophysiology. In this report, we focused on using Wnt inhibitors to study the differentiation of human iPSC-CPCs under 2D or 3D culture conditions by measuring marker protein and gene expression as well as intracellular Ca2+ oscillation. Our results show that the 3D culture with aligned nanofiber scaffolds, mimicing the architecture of the extracellular matrix of the heart, improve the differentiation of iPSC-CPCs to functional cardiomyocytes induced by Wnt inhibition, as shown with increased number of cardiac Troponin T (cTnT)-positive cells and synchronized intracellular Ca2+ oscillation. In addition, we studied if 3D nanofiber culture can be used as an in vitro model for compound screening by testing a number of other differentiation factors including a ALK5 inhibitor and inhibitors of BMP signaling. This work highlights the importance of using a more relevant in vitro model and measuring not only the expression of marker proteins but also the functional readout in a screen in order to identify the best compounds and to investigate the resulting biology.