Study of myoblast differentiation using multi-dimensional scaffolds consisting of nano and micropatterns.
Study of myoblast differentiation using multi-dimensional scaffolds consisting of nano and micropatterns.
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DOI：
10.1186/s40824-016-0087-x
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发表时间：
2017
影响因子：
11.3
通讯作者：
Koh WG
中科院分区：
文献类型：
--
作者：
Cha SH;Lee HJ;Koh WG
地形线索是骨骼肌细胞培养的主要影响，因为结构是高度组织化的，由成肌细胞卫星细胞分化和融合形成的多核肌管的长平行束组成。在这份技术报告中，我们使用静电纺丝和聚乙二醇（PEG）水凝胶微图案制造了一个多尺度支架，以监测具有不同角度组合的纳米和微米排列组合支架上的细胞行为。我们制造了多尺度支架，通过静电纺丝和PEG水凝胶微图案化提供生物相容性和细胞外基质（ECM）模拟环境。MTT分析表明，在7天的细胞培养期间，所有实验组的增殖率几乎增加了四倍。测量细胞取向和伸长以确认成肌潜力。在对齐的纤维支架上，超过90%的细胞分散在纤维方向的± 20°。为了确定细胞伸长，我们监测了细胞核的纵横比。在无规纤维上，细胞表现出1.33的纵横比，但在垂直和平行纳米纤维上，纵横比大于2。肌球蛋白重链（MHC）表达显著高于i）平行纤维与随机纤维相比，ii）100 μm与200 μm线状模式相比。我们证实了肌管形成的不同趋势，可以通过多维支架激发。我们研究了更有利的环境，通过结合纳米和微米尺度的模式，诱导细胞排列和延长肌生成。该系统可以作为一种新型的多维平台来研究体外细胞行为。
The topographical cue is major influence on skeletal muscle cell culture because the structure is highly organized and consists of long parallel bundles of multinucleated myotubes that are formed by differentiation and fusion of myoblast satellite cells. In this technical report, we fabricated a multiscale scaffold using electrospinning and poly (ethylene glycol) (PEG) hydrogel micropatterns to monitor the cell behaviors on nano- and micro-alignment combined scaffolds with different combinations of angles. We fabricated multiscale scaffolds that provide biocompatible and extracellular matrix (ECM)-mimetic environments via electrospun nanofiber and PEG hydrogel micro patterning. MTT assays demonstrated an almost four-fold increase in the proliferation rate during the 7 days of cell culture for all of the experimental groups. Cell orientation and elongation were measured to confirm the myogenic potential. On the aligned fibrous scaffolds, more than 90% of the cells were dispersed ± 20° of the fiber orientation. To determine cell elongation, we monitored nuclei aspect ratios. On a random nanofiber, the cells demonstrated an aspect ratio of 1.33, but on perpendicular and parallel nanofibers, the aspect ratio was greater than 2. Myosin heavy chain (MHC) expression was significantly higher i) on parallel compared to random fibers, ii) the 100 μm compared to the 200 μm line pattern. We confirmed the disparate trends of myotube formation that can be provoked through multi-dimensional scaffolds. We studied more favorable environments that induce cell alignment and elongation for myogenesis by combining nano- and micro-scale patterns. The fabricated system can serve as a novel multi-dimensional platform to study in vitro cell behaviors.