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首页> 外文期刊>Skeletal Muscle >Culturing C2C12 myotubes on micromolded gelatin hydrogels accelerates myotube maturation
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Culturing C2C12 myotubes on micromolded gelatin hydrogels accelerates myotube maturation

机译：培养C2C12 Microded Gelatin水凝胶的Myotubes加速MyOTube成熟

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Skeletal muscle contributes to roughly 40% of lean body mass, and its loss contributes to morbidity and mortality in a variety of pathogenic conditions. Significant insights into muscle function have been made using cultured cells, in particular, the C2C12 myoblast line. However, differentiation of these cells in vitro typically yields immature myotubes relative to skeletal muscles in vivo. While many efforts have attempted to improve the maturity of cultured myotubes, including the use of bioengineered substrates, lack of molecular characterization has precluded their widespread implementation. This study characterizes morphological, molecular, and transcriptional features of C2C12 myotubes cultured on crosslinked, micropatterned gelatin substrates fabricated using previously established methods and compares them to myotubes grown on unpatterned gelatin or traditional plasticware. We used immunocytochemistry, SDS-PAGE, and RNAseq to characterize C2C12 myotubes grown on micropatterned gelatin hydrogels, unpatterned gelatin hydrogels, and typical cell culture substrates (i.e., plastic or collagen-coated glass) across a differentiation time course. The ability to form aligned sarcomeres and myofilament protein concentration was assessed. Additionally, the transcriptome was analyzed across the differentiation time course. C2C12 myotubes grown on micropatterned gelatin hydrogels display an increased ability to form aligned sarcomeres as well as increased contractile protein content relative to myotubes cultured on unpatterned gelatin and plastic. Additionally, genes related to sarcomere formation and in vivo muscle maturation are upregulated in myotubes grown on micropatterned gelatin hydrogels relative to control myotubes. Our results suggest that growing C2C12 myotubes on micropatterned gelatin hydrogels accelerates sarcomere formation and yields a more fully matured myotube culture. Thus, the use of micropatterned hydrogels is a viable and simple approach to better model skeletal muscle biology in vitro.
机译：骨骼肌占瘦体重的约40％，其损失有助于各种致病病症的发病率和死亡率。使用培养的细胞，特别是C2C12肌细胞线进行了对肌肉功能的显着洞察。然而，在体外的这些细胞的分化通常通常相对于体内骨骼肌产生不成熟的肌管。虽然许多努力已经尝试改善培养的肌管的成熟，但包括使用生物工程底物的使用，缺乏分子表征妨碍了它们广泛的实施。本研究表征了使用先前建立的方法培养的C2C12 Myotubes的形态学，分子和转录特征，并将其与在未绘图的明胶或传统塑料中生长的肌管进行比较。我们使用免疫细胞化学，SDS-PAGE和RNASEQ在微过图案化明胶水凝胶，未绘图的明胶水凝胶上生长的C2C12 Myotubes，以及穿过分化时间过程的典型细胞培养物和典型的细胞培养基材（即塑料或胶原涂层玻璃）。评估形成对齐的SARCOMERS和硫丝蛋白浓度的能力。另外，在分化时间过程中分析转录组。 C2C12在微透明的明胶水凝胶上生长的Myotubes显示出形成对齐的SARCOMERES的能力，以及相对于在未绘图的明胶和塑料上培养的肌管的增加的收缩蛋白质含量。另外，与Sarcomere形成和体内肌肉成熟有关的基因在微透过的明胶水凝胶中生长的肌管相对于对照肌管中的肌管上调。我们的研究结果表明，在微明犹太明明胶水凝胶上生长C2C12肌管加速了SARCOME系的形成，并产生了更完全成熟的肌管培养物。因此，使用微图案化水凝胶是一种可行且简单的方法，可以在体外更好地模型骨骼肌生物学。

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