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首页> 外文期刊>Acta biomaterialia >Long-term culture of HL-1 cardiomyocytes in modular poly(ethylene glycol) microsphere-based scaffolds crosslinked in the phase-separated state
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Long-term culture of HL-1 cardiomyocytes in modular poly(ethylene glycol) microsphere-based scaffolds crosslinked in the phase-separated state

机译：HL-1心肌细胞在相分离状态下交联的模块化聚乙二醇微球基支架中的长期培养

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摘要

Poly(ethylene glycol) (PEG) microspheres were assembled around HL-1 cardiomyocytes to produce highly porous modular scaffolds. In this study we took advantage of the immiscibility of PEG and dextran to improve upon our previously described modular scaffold fabrication methods. Phase separating the PEG microspheres in dextran solutions caused them to rapidly deswell and crosslink together, eliminating the need for serum protein-based crosslinking. This also led to a dramatic increase in the stiffness of the scaffolds and greatly improved the handling characteristics. HL-1 cardiomyocytes were present during microsphere crosslinking in the cytocompatible dextran solution, exhibiting high cell viability following scaffold formation. Over the course of 2 weeks a 9-fold expansion in cell number was observed. The cardiac functional markers sarcomeric α-actinin and connexin 43 were expressed at 13 and 24 days after scaffold formation. HL-1 cells were spontaneously depolarizing 38 days after scaffold formation, which was visualized by confocal microscopy using a calcium-sensitive dye. Electrical stimulation resulted in synchronization of activation peaks throughout the scaffolds. These findings demonstrate that PEG microsphere scaffolds fabricated in the presence of dextran can support the long-term three-dimensional culture of cells, suggesting applications in cardiovascular tissue engineering.
机译：聚（乙二醇）（PEG）微球在HL-1心肌细胞周围组装，以生产高度多孔的模块化支架。在这项研究中，我们利用了PEG和葡聚糖的不混溶性来改进我们先前描述的模块化支架制造方法。葡聚糖溶液中PEG微球的相分离使它们迅速溶胀并交联在一起，从而消除了基于血清蛋白的交联的需要。这也导致了支架的刚度的显着提高，并大大改善了操作特性。 HL-1心肌细胞在微球交联过程中存在于细胞相容性葡聚糖溶液中，在形成支架后表现出高细胞活力。在2周的过程中，观察到细胞数扩展了9倍。心脏功能标记肌节α-肌动蛋白和连接蛋白43在支架形成后第13和24天表达。 HL-1细胞在支架形成后38天自发去极化，这可通过使用钙敏感染料的共聚焦显微镜观察。电刺激导致整个支架的激活峰同步。这些发现表明，在葡聚糖的存在下制备的PEG微球支架可以支持细胞的长期三维培养，表明其在心血管组织工程中的应用。

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