Antin P B, Ordahl C P
Department of Anatomy, University of California, San Francisco 94143.
Dev Biol. 1991 Jan;143(1):111-21. doi: 10.1016/0012-1606(91)90058-b.
Myogenic cell lines have proven extremely valuable for studying myogenesis in vitro. Although a number of mammalian muscle cell lines have been isolated, attempts to produce cell lines from other classes of animals have met with only limited success. We report here the isolation and characterization of seven avian myogenic cell lines (QM1-4 and QM6-8), derived from the quail fibrosarcoma cell line QT6. A differentiation incompetent QM cell derivative was also isolated (QM5DI). The major features of QM cell differentiation in vitro closely resemble those of their mammalian counterparts. Mononucleated QM cells replicate in medium containing high concentrations of serum components. Upon switching to medium containing low serum components, cells withdraw from the cell cycle and fuse to form elongated multinucleated myotubes. Cultures typically obtain fusion indices of 43-49%. Northern blot and immunoblot analyses demonstrate that each differentiated QM cell line expresses a wide variety of genes encoding muscle specific proteins: desmin, cardiac troponin T, skeletal troponin T, cardiac troponin C, skeletal troponin I, alpha-tropomyosin, muscle creatine kinase, myosin light chain 2, and a ventricular isoform of myosin heavy chain. While all QM lines analyzed to date express at least some myosin light chain 2, only one line, QM7, expresses this gene at high levels. Surprisingly, none of the QM lines reported here express any known form of alpha-actin. The absence of sarcomeric actin expression may explain the absence of myofibrils in QM myotubes. These novel features of muscle gene expression in QM cells may prove useful for studying the role of specific muscle proteins during myogenesis. More importantly, however, the isolation of QM cell lines indicates that it may be feasible to isolate other avian myogenic cell lines with general utility for the study of muscle development.
肌源性细胞系已被证明在体外研究肌生成方面极具价值。尽管已经分离出了许多哺乳动物肌肉细胞系,但从其他动物类别中产生细胞系的尝试仅取得了有限的成功。我们在此报告从鹌鹑纤维肉瘤细胞系QT6中分离并鉴定出七个禽类肌源性细胞系(QM1 - 4和QM6 - 8)。还分离出了一种无分化能力的QM细胞衍生物(QM5DI)。QM细胞在体外的主要分化特征与它们的哺乳动物对应物非常相似。单核的QM细胞在含有高浓度血清成分的培养基中复制。当转换到含有低血清成分的培养基时,细胞退出细胞周期并融合形成细长的多核肌管。培养物通常获得43 - 49%的融合指数。Northern印迹和免疫印迹分析表明,每个分化的QM细胞系都表达多种编码肌肉特异性蛋白质的基因:结蛋白、心肌肌钙蛋白T、骨骼肌肌钙蛋白T、心肌肌钙蛋白C、骨骼肌肌钙蛋白I、α - 原肌球蛋白、肌肉肌酸激酶、肌球蛋白轻链2以及肌球蛋白重链的心室同工型。虽然迄今为止分析的所有QM系都至少表达一些肌球蛋白轻链2,但只有一个系QM7高水平表达该基因。令人惊讶的是,这里报道的QM系均不表达任何已知形式的α - 肌动蛋白。肌节肌动蛋白表达的缺失可能解释了QM肌管中肌原纤维的缺失。QM细胞中肌肉基因表达的这些新特征可能有助于研究特定肌肉蛋白在肌生成过程中的作用。然而,更重要的是,QM细胞系的分离表明分离其他具有普遍用途以研究肌肉发育的禽类肌源性细胞系可能是可行的。