Zhang Mingming, Xu Yuerong, Chen Yan, Yan Qinru, Li Xiaoli, Ding Lu, Wei Ting, Zeng Di
Department of Cardiology, Tangdu Hospital, the Fourth Military Medical University, Xi'an, China.
Department of Orthodontics, School of Stomatology, the Fourth Military Medical University, Xi'an, China.
Front Cell Dev Biol. 2022 Aug 1;10:875278. doi: 10.3389/fcell.2022.875278. eCollection 2022.
Although pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) have been proved to be a new platform for heart regeneration, the lack of maturity significantly hinders the clinic application. Recent researches indicate that the function of stem cell is associated with the nanoscale geometry/topography of the extracellular matrix (ECM). However, the effects of 3D nanofibrous scaffolds in maturation of iPSC-CMs still remain unclear. Thus, we explored the effects of restructuring iPSC-CMs in 3D nano-scaffolds on cell morphology, cardiac-specific structural protein, gap junction and calcium transient kinetics. Using the electrospinning technology, poly-(ε-caprolactone) (PCL) nanofibrous scaffold were constructed and iPSC-CMs were seeded into these forms. As expected, strong sarcolemmal remodeling processes and myofilament reorientation were observed in 3D nano-scaffolds culture, as well as more expression of cardiac mature proteins, such as β-MHC and MLC2v. The mature morphology of 3D-shaped iPSC-CMs leaded to enhanced calcium transient kinetics, with increased calcium peak transient amplitude and the maximum upstroke velocity (Vmax). The results revealed that the maturation of iPSC-CMs was enhanced by the electrospun 3D PCL nanofibrous scaffolds treatment. These findings also proposed a feasible strategy to improve the myocardium bioengineering by combining stem cells with scaffolds.
尽管多能干细胞衍生的心肌细胞（iPSC-CMs）已被证明是心脏再生的新平台，但缺乏成熟度严重阻碍了其临床应用。最近的研究表明，干细胞的功能与细胞外基质（ECM）的纳米级几何形状/拓扑结构有关。然而，3D纳米纤维支架对iPSC-CMs成熟的影响仍不清楚。因此，我们探讨了在3D纳米支架中重组iPSC-CMs对细胞形态、心脏特异性结构蛋白、间隙连接和钙瞬变动力学的影响。利用静电纺丝技术构建了聚（ε-己内酯）（PCL）纳米纤维支架，并将iPSC-CMs接种到这些支架中。正如预期的那样，在3D纳米支架培养中观察到了强烈的肌膜重塑过程和肌丝重新定向，以及心脏成熟蛋白如β-MHC和MLC2v的更多表达。3D形状的iPSC-CMs的成熟形态导致钙瞬变动力学增强，钙瞬变峰值幅度和最大上升速度（Vmax）增加。结果表明，静电纺丝3D PCL纳米纤维支架处理可增强iPSC-CMs的成熟度。这些发现还提出了一种通过将干细胞与支架相结合来改善心肌生物工程的可行策略。
