Rhim Caroline, Lowell Dorothy A, Reedy Mary C, Slentz Dorothy H, Zhang Sarah J, Kraus William E, Truskey George A
Department of Biomedical Engineering, Duke University, 136 Hudson Hall, Box 90281, Durham, North Carolina 27708, USA.
Muscle Nerve. 2007 Jul;36(1):71-80. doi: 10.1002/mus.20788.
Because previous studies of three-dimensional skeletal muscle cultures have shown limited differentiation, the goal of this study was to establish conditions that would produce mature sarcomeres in a mammalian-derived skeletal muscle construct. We evaluated the differentiation of bioartificial muscles generated from C(2)C(12) myoblasts in a collagen gel cultured under steady, passive tension for up to 36 days. Staining for alpha-actinin, myosin, and F-actin indicated the presence of striated fibers as early as 6 days post-differentiation. Electron microscopy at 16 days post-differentiation revealed multinucleated myotubes with ordered, striated myofibers. At 33 days, the cultures contained collagen fibers and showed localization of paxillin at the fiber termini, suggesting that myotendinous junctions were forming. The present study demonstrates mature muscle synthesis in a three-dimensional system using a pure mammalian myoblast cell line. Our results suggest that this culture model can be used to evaluate the effects of various mechanical and biochemical cues on muscle development under normal and pathological conditions.
由于先前对三维骨骼肌培养的研究显示分化有限，本研究的目标是建立能在源自哺乳动物的骨骼肌构建体中产生成熟肌节的条件。我们评估了在稳定、被动张力下培养长达36天的胶原蛋白凝胶中由C(2)C(12)成肌细胞生成的生物人工肌肉的分化情况。α-辅肌动蛋白、肌球蛋白和F-肌动蛋白染色表明，早在分化后6天就存在横纹肌纤维。分化后16天的电子显微镜检查显示有多核肌管，其肌纤维有序且有横纹。在33天时，培养物中含有胶原纤维，并且桩蛋白定位于纤维末端，表明肌-腱连接正在形成。本研究证明了使用纯哺乳动物成肌细胞系在三维系统中合成成熟肌肉。我们的结果表明，这种培养模型可用于评估在正常和病理条件下各种机械和生化信号对肌肉发育的影响。
