Understanding the Role of ECM Protein Composition and Geometric Micropatterning for Engineering Human Skeletal Muscle.
Understanding the Role of ECM Protein Composition and Geometric Micropatterning for Engineering Human Skeletal Muscle.
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DOI：
10.1007/s10439-016-1592-8
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发表时间：
2016-06
影响因子：
3.8
通讯作者：
Feinberg AW
中科院分区：
文献类型：
--
作者：
Duffy RM;Sun Y;Feinberg AW
由于将人成肌细胞分化成对齐的可收缩组织的挑战，已经证明通过创伤或疾病失去的骨骼肌难以再生。为了解决这个问题，我们研究了微环境的线索，驱动成肌细胞分化成对齐的肌管在骨骼肌修复，器官芯片疾病模型和软机器人致动器的潜在应用。我们使用2D体外系统来系统地评估细胞外基质（ECM）蛋白质组成和几何图案对控制高度对齐的肌管形成的作用。具体而言，我们分析了分化的肌管从鼠C2C12细胞和人骨骼肌衍生细胞（SkMDC）的层粘连蛋白相比，纤连蛋白，胶原蛋白I型，和胶原蛋白IV型的微图案线。结果显示，层粘连蛋白支持两种细胞类型的显著更大的肌管形成，导致与其他ECM蛋白相比，这些表面上的肌管面积增加超过2倍。物种特异性差异显示，人类SkMDC在广泛的微图案化线条尺寸上单轴对齐，而C2C12需要特定的线条宽度和间距才能做到这一点。未来的工作将结合这些结果，在2D中设计对齐的人类骨骼肌组织，用于疾病建模，药物发现和毒性筛选的体外应用。
Skeletal muscle lost through trauma or disease has proven difficult to regenerate due to the challenge of differentiating human myoblasts into aligned, contractile tissue. To address this, we investigated microenvironmental cues that drive myoblast differentiation into aligned myotubes for potential applications in skeletal muscle repair, organ-on-chip disease models and actuators for soft robotics. We used a 2D in vitro system to systematically evaluate the role of extracellular matrix (ECM) protein composition and geometric patterning for controlling the formation of highly aligned myotubes. Specifically, we analyzed myotubes differentiated from murine C2C12 cells and human skeletal muscle derived cells (SkMDCs) on micropatterned lines of laminin compared to fibronectin, collagen type I, and collagen type IV. Results showed that laminin supported significantly greater myotube formation from both cells types, resulting in greater than 2-fold increase in myotube area on these surfaces compared to the other ECM proteins. Species specific differences revealed that human SkMDCs uniaxially aligned over a wide range of micropatterned line dimensions, while C2C12s required specific line widths and spacings to do the same. Future work will incorporate these results to engineer aligned human skeletal muscle tissue in 2D for in vitro applications in disease modeling, drug discovery and toxicity screening.