Differentiation of myoblasts in serum-free media: Effects of modified media are cell line-specific
Differentiation of myoblasts in serum-free media: Effects of modified media are cell line-specific
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DOI：
10.1159/000016776
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发表时间：
2000-01-01
影响因子：
2.7
通讯作者：
Purslow, PP
中科院分区：
文献类型：
--
作者：
Lawson, MA;Purslow, PP
成肌细胞系在细胞培养物中容易生长和分化。通常用于研究肌肉生长和分化的两种细胞系是小鼠细胞系C2C12和大鼠细胞系L6。这些细胞的体外分化需要在细胞达到汇合后从富含血清的培养基转换到不太丰富的培养基。由于血清中存在的组分尚未得到充分表征，因此研究了使用更好的成分确定培养基进行这些研究。C2C12和L6成肌细胞在含血清和无血清培养基中分化。然后用显微镜和生化方法检测这些培养物的分化状态。检查培养物的肌管形成、肌酸磷酸激酶的活性和肌节肌动蛋白的存在。在C2C12鳗鱼Is中，无血清系统的分化程度大于含血清系统。在两种培养基中，C2C12细胞产生肌节肌动蛋白，表明肌管中存在肌节结构。然而，在L6细胞中，肌管在含有2%马血清的培养基中容易形成，但在无血清系统中不容易形成。此外，C2C12细胞在包被有细胞外基质蛋白的基质上分化的能力被证明是介质依赖性的。细胞外基质蛋白的存在并不能使L6细胞在无血清培养基中培养时形成肌管。鸡成肌细胞的原代培养物能够在两种培养基中分化，含有马血清的Dulbecco改良Eagle培养基是细胞融合的更有效培养基。这项研究显示了三种细胞系中肌肉细胞系反应的差异，其中两种通常用作理解肌肉生长和发育的“模型系统”。版权所有（C）2000 S. Karger AG，巴塞尔。
Myoblast cell lines are grown and differentiated readily in cell culture. Two cell lines typically used for investigating the growth and differentiation of muscle are the mouse cell line C2C12 and the rat cell line L6. The differentiation of these cells in vitro requires a switch from a serum-rich medium to a less rich medium after the cells have reached confluence. Since the components present in serum are not well characterized, the use of a better defined medium for these studies was investigated. C2C12 and L6 myoblasts were differentiated in both serum-containing and serum-free media. The differentiation state of these cultures was then tested both microscopically and biochemically. Cultures were checked for myotube formation, the activity of creatine phosphokinase and the presence of sarcomeric actin. In C2C12 eel Is, the extent of differentiation was greater in the serum-free than in the serum-containing system. in both media types, the C2C12 cells produced sarcomeric actin, showing the presence of sarcomere structure in the myotubes. In L6 cells, however, myotubes were readily formed in medium containing 2% horse serum, but not in the serum-free system. In addition, the ability of C2C12 cells to differentiate on substrates coated with extracellular matrix proteins was shown to be media-dependent. The presence of extracellular matrix proteins did not enable L6 cells to form myotubes when cultured in serum-free media. Primary cultures of chick myoblasts were able to differentiate in both media tested, with Dulbecco's modified Eagle medium containing horse serum being a more efficient medium for cell fusion. This study shows a divergence in muscle cell line responses in th ree cell lines, two of which are typically used as 'model systems' for understanding muscle growth and development. Copyright (C) 2000 S. Karger AG,Basel.