Temporal impact of substrate mechanics on differentiation of human embryonic stem cells to cardiomyocytes.
Temporal impact of substrate mechanics on differentiation of human embryonic stem cells to cardiomyocytes.
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DOI：
10.1016/j.actbio.2013.10.033
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发表时间：
2014-02
影响因子：
9.7
通讯作者：
Palecek, Sean P.
中科院分区：
文献类型：
--
作者：
Hazeltine, Laurie B.;Badur, Mehmet G.;Lian, Xiaojun;Das, Amritava;Han, Wenqing;Palecek, Sean P.
将人类多能干细胞（hPSCs）分化为心肌细胞是一项重要的临床需求，为体内心脏修复的新药物或细胞治疗的体外组织建模提供了可能。众所周知，化学和机械微环境因素会影响干细胞分化的效率，但hPSCs的心脏分化方案通常在刚性组织培养聚苯乙烯（TCPS）表面进行，不存在生理机械设置。为了研究力学对心脏分化的时间效应，我们将人胚胎干细胞（hESCs）及其衍生物培养在具有生理相关刚度范围的聚丙烯酰胺水凝胶基质上。在定向分化和胚状体培养系统中，hESCs向表达心肌肌钙蛋白t （cTnT+）的心肌细胞的分化在中等硬度的水凝胶上达到顶峰。在定向分化的第1天，Brachyury在中等硬度水凝胶上的表达也达到峰值，表明硬度影响了hESCs向中胚层的初始分化轨迹。为了研究底物力学在中胚层祖细胞心脏分化过程中的影响，我们在Nkx2.5/Isl1+心脏祖细胞阶段在TCPS上启动了定向心肌细胞分化，并将细胞转移到水凝胶中。第15天心肌细胞纯度与僵硬度无差异。这些实验表明，在中胚层诱导的早期阶段，hESCs的分化对底物力学敏感，适当应用底物力学可以增加hESCs向心肌细胞分化的倾向。
A significant clinical need exists to differentiate human pluripotent stem cells (hPSCs) into cardiomyocytes, enabling tissue modeling for in vitro discovery of new drugs or cell-based therapies for heart repair in vivo. Chemical and mechanical microenvironmental factors are known to impact efficiency of stem cell differentiation, but cardiac differentiation protocols in hPSCs are typically performed on rigid tissue culture polystyrene (TCPS) surfaces which do not present a physiological mechanical setting. To investigate the temporal effects of mechanics on cardiac differentiation, we cultured human embryonic stem cells (hESCs) and their derivatives on polyacrylamide hydrogel substrates with a physiologically relevant range of stiffnesses. In directed differentiation and embryoid body culture systems, differentiation of hESCs to cardiac Troponin T-expressing (cTnT+) cardiomyocytes peaked on hydrogels of intermediate stiffness. Brachyury expression also peaked on intermediate stiffness hydrogels at day 1 of directed differentiation, suggesting that stiffness impacted the initial differentiation trajectory of hESCs to mesendoderm. To investigate the impact of substrate mechanics during cardiac specification of mesodermal progenitors, we initiated directed cardiomyocyte differentiation on TCPS and transferred cells to hydrogels at the Nkx2.5/Isl1+ cardiac progenitor cell stage. No differences in cardiomyocyte purity with stiffness were observed on day 15. These experiments indicate that differentiation of hESCs is sensitive to substrate mechanics at early stages of mesodermal induction, and proper application of substrate mechanics can increase the propensity of hESCs to differentiate to cardiomyocytes.
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影响因子：
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作者：
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