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Prolonged Culture of Aligned Skeletal Myotubes on Micromolded Gelatin Hydrogels

机译：微胶质明胶水凝胶的延长培养物对齐骨髓肌管

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In vitro models of skeletal muscle are critically needed to elucidate disease mechanisms, identify therapeutic targets, and test drugs pre-clinically. However, culturing skeletal muscle has been challenging due to myotube delamination from synthetic culture substrates approximately one week after initiating differentiation from myoblasts. In this study, we successfully maintained aligned skeletal myotubes differentiated from C2C12 mouse skeletal myoblasts for three weeks by utilizing micromolded (μmolded) gelatin hydrogels as culture substrates, which we thoroughly characterized using atomic force microscopy (AFM). Compared to polydimethylsiloxane (PDMS) microcontact printed (μprinted) with fibronectin (FN), cell adhesion on gelatin hydrogel constructs was significantly higher one week and three weeks after initiating differentiation. Delamination from FN-μprinted PDMS precluded robust detection of myotubes. Compared to a softer blend of PDMS μprinted with FN, myogenic index, myotube width, and myotube length on μmolded gelatin hydrogels was similar one week after initiating differentiation. However, three weeks after initiating differentiation, these parameters were significantly higher on μmolded gelatin hydrogels compared to FN-μprinted soft PDMS constructs. Similar results were observed on isotropic versions of each substrate, suggesting that these findings are independent of substrate patterning. Our platform enables novel studies into skeletal muscle development and disease and chronic drug testing in vitro.
机译：骨骼肌的体外模型主要需要阐明疾病机制，临床鉴定治疗靶标和测试药物。然而，在从肌细胞分化的分化后大约一周，培养骨骼肌已经挑战，由于肌肌细胞肌肌法从肌细胞分化约一周。在这项研究中，我们通过利用微胶质（μmolded）明胶水凝胶作为培养基底物，成功地保持了与C2C12小鼠骨骨菌丝细胞的对齐骨髓肌细胞三周。我们用原子力显微镜（AFM）彻底表征。与聚二甲基硅氧烷（PDMS）微接触（μPrexted）与纤连蛋白（Fn）相比，在引发分化后明胶水凝胶构建体上的细胞粘附明显高出3周。来自FN-μprexed的PDMS的分层排除了肌管的鲁棒检测。与具有Fn的PDMSμprexed的较软混合物相比，在引发分化后一周内与μmoldedGelatin水凝胶上的肌原素指数，肌室宽度和肌室长度相似。然而，与Fn-μprepted软PDMS构建体相比，在引发分化后三周后，这些参数显着高于μmmoldedGelatin水凝胶。在每个基材的各向同性型号上观察到类似的结果，表明这些发现与基板图案相比。我们的平台使新的研究能够进入骨骼肌发育和疾病和体外慢性药物检测。

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