Engineering Muscle Networks in 3D Gelatin Methacryloyl Hydrogels: Influence of Mechanical Stiffness and Geometrical Confinement.
Engineering Muscle Networks in 3D Gelatin Methacryloyl Hydrogels: Influence of Mechanical Stiffness and Geometrical Confinement.
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DOI：
10.3389/fbioe.2017.00022
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发表时间：
2017
影响因子：
5.7
通讯作者：
Rainer A
中科院分区：
文献类型：
--
作者：
Costantini M;Testa S;Fornetti E;Barbetta A;Trombetta M;Cannata SM;Gargioli C;Rainer A
在这项工作中，从体外肌生成的角度评估了机械刚度和几何约束对成肌细胞负载的明胶甲基丙烯酰 (GelMA) 光交联水凝胶 3D 培养的影响。我们配制了一组充满细胞的 GelMA 水凝胶，其压缩模量在 1 ÷17kPa 范围内，通过改变 GelMA 浓度和交联程度获得。选择 C2C12 成肌细胞作为细胞模型，研究不同 GelMA 水凝胶对肌管形成长达 2 周的支持作用。结果表明，硬度在 1 ÷3 kPa 范围内的水凝胶在肌管数量、形成速率和空间分布方面为 C2C12 分化提供了增强的支持。最后，我们通过将细胞限制在具有不同横截面的薄水凝胶板内，研究了几何限制对肌管方向的影响：（i）2,000 μm × 2,000 μm，（ii）1,000 μm × 1,000 μm，（iii） 500 μm × 500 μm。获得的结果表明，通过减少横截面，即通过增加限制水平，肌管更加紧密地堆积并形成更好地模仿骨骼肌天然形态的对齐肌结构。
In this work, the influence of mechanical stiffness and geometrical confinement on the 3D culture of myoblast-laden gelatin methacryloyl (GelMA) photo-crosslinkable hydrogels was evaluated in terms of in vitro myogenesis. We formulated a set of cell-laden GelMA hydrogels with a compressive modulus in the range 1 ÷ 17 kPa, obtained by varying GelMA concentration and degree of cross-linking. C2C12 myoblasts were chosen as the cell model to investigate the supportiveness of different GelMA hydrogels toward myotube formation up to 2 weeks. Results showed that the hydrogels with a stiffness in the range 1 ÷ 3 kPa provided enhanced support to C2C12 differentiation in terms of myotube number, rate of formation, and space distribution. Finally, we studied the influence of geometrical confinement on myotube orientation by confining cells within thin hydrogel slabs having different cross sections: (i) 2,000 μm × 2,000 μm, (ii) 1,000 μm × 1,000 μm, and (iii) 500 μm × 500 μm. The obtained results showed that by reducing the cross section, i.e., by increasing the level of confinement—myotubes were more closely packed and formed aligned myostructures that better mimicked the native morphology of skeletal muscle.