Regulating Fibrinolysis to Engineer Skeletal Muscle from the C2C12 Cell Line
Regulating Fibrinolysis to Engineer Skeletal Muscle from the C2C12 Cell Line
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DOI：
10.1089/ten.tec.2008.0286
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发表时间：
2009-09-01
影响因子：
3
通讯作者：
Baar, Keith
中科院分区：
文献类型：
--
作者：
Khodabukus, Alastair;Baar, Keith
从转化细胞工程化的肌肉将是一个强大的模型，通过允许长期的肌肉适应体外研究的肌肉生理学研究。然而，先前描述的方法要么花费>5周来产生组织，要么使用胶原蛋白作为支架，这降低了肌肉的比力，使得难以测量构建体的功能。本研究的目的是使用纤维蛋白中的C2 C12细胞系快速工程化肌肉，纤维蛋白具有与肌肉组织相似的硬度，允许准确的功能测试。蛋白酶抑制剂抑肽酶和天然交联剂京尼平都增加了肌肉可以培养的时间长度，京尼平将培养时间增加到10周。组织的功能受到血清（64-78%）或凝血酶（41%）批次、分化培养基（78%）和接种方案（38%）的显著影响，但不受初始细胞数的影响。引人注目的是，不同的C2 C12克隆产生高达3.6倍的力产生变化。在最佳条件下，组织在10.4 +/- 0.3天内形成，并在5周内保持完全功能，在此期间它们继续成熟。这里描述的优化模型提供了快速，可靠和功能性组织，这将是有用的骨骼肌生理学的研究。
Muscles engineered from transformed cells would be a powerful model for the study of muscle physiology by allowing long-term in vitro studies of muscle adaptation. However, previously described methods either take >5 weeks to produce a tissue or use collagen as a scaffold, which decreases the specific force of the muscle, making it hard to measure the function of the constructs. The aim of this study was to rapidly engineer muscle using the C2C12 cell line in fibrin, which has a stiffness similar to muscle tissue, allowing accurate functional testing. Both the protease inhibitor aprotinin and the natural cross-linker genipin increased the length of time that muscle could be cultured, with genipin increasing the time in culture to 10 weeks. The function of the tissues was significantly affected by the batch of serum (64-78%) or thrombin (41%), the differentiation medium (78%), and the seeding protocol (38%), but was unaffected by initial cell number. Strikingly, different C2C12 clones produced up to a 3.6-fold variation in force production. Under optimal conditions, the tissues form in 10.4 +/- 0.3 days and remain fully functional for 5 weeks over which time they continue to mature. The optimized model described here provides rapid, reliable, and functional tissues that will be useful in the study of skeletal muscle physiology.