Cell alignment induced by anisotropic electrospun fibrous scaffolds alone has limited effect on cardiomyocyte maturation.
Cell alignment induced by anisotropic electrospun fibrous scaffolds alone has limited effect on cardiomyocyte maturation.
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DOI：
10.1016/j.scr.2016.04.014
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发表时间：
2016-05
影响因子：
1.2
通讯作者：
Xu C
中科院分区：
文献类型：
--
作者：
Han J;Wu Q;Xia Y;Wagner MB;Xu C
促进人多能干细胞衍生的心肌细胞（hPSC-CM）的成熟将促进其在疾病建模和药物发现中的应用。以前的研究表明，细胞对齐可以提高hPSC-CM成熟，然而，这种方法的鲁棒性还没有得到很好的研究。为此，我们检查了由3D微环境内的底层对齐纤维施加的hPSC-CM的各向异性取向是否可以改善hPSC-CM的成熟。将富集的hPSC-CM在基质胶涂覆的各向异性（对齐的）和各向同性（随机的）聚己内酯（PCL）纤维支架上培养两周，以及组织培养聚苯乙烯（TCP）作为对照。正如预期的那样，在两种类型的纤维支架上生长的hPSC-CM分别表现出各向异性和各向同性取向。与TCP上的细胞类似，在这些支架上培养的hPSC-CM表达CM相关蛋白，并且以剂量依赖性方式对肾上腺素能受体激动剂、毒蕈碱激动剂和间隙连接解偶联剂产生强烈反应。虽然生长在各向异性纤维支架上的hPSC-CM显示出编码许多肌节蛋白、钙处理蛋白和离子通道的基因的最高表达，但它们的钙瞬时动力学比生长在TCP上的细胞慢。这些结果表明，电纺各向异性纤维支架，作为一种单一的方法，对提高hPSC-CM的成熟的效果有限。
Enhancing the maturation of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) will facilitate their applications in disease modeling and drug discovery. Previous studies suggest that cell alignment could enhance hPSC-CM maturation; however, the robustness of this approach has not been well investigated. To this end, we examined if the anisotropic orientation of hPSC-CMs imposed by the underlying aligned fibers within a 3D microenvironment could improve the maturation of hPSC-CMs. Enriched hPSC-CMs were cultured for two weeks on Matrigel-coated anisotropic (aligned) and isotropic (random) polycaprolactone (PCL) fibrous scaffolds, as well as tissue culture polystyrenes (TCPs) as a control. As expected, hPSC-CMs grown on the two types of fibrous scaffolds exhibited anisotropic and isotropic orientations, respectively. Similar to cells on TCPs, hPSC-CMs cultured on these scaffolds expressed CM-associated proteins and were pharmacologically responsive to adrenergic receptor agonists, a muscarinic agonist, and a gap junction uncoupler in a dose-dependent manner. Although hPSC-CMs grown on anisotropic fibrous scaffolds displayed the highest expression of genes encoding a number of sarcomere proteins, calcium handling proteins and ion channels, their calcium transient kinetics were slower than cells grown on TCPs. These results suggest that electrospun anisotropic fibrous scaffolds, as a single method, have limited effect on improving the maturation of hPSC-CMs.