The availability of an in vitro canine cell line would reduce the need for dogs for primary in vitro cell culture and reduce overall cost in pre-clinical studies. An immortalized canine muscle cell line, named Myok9, from primary myoblasts of a normal dog has been developed by the authors. Immortalization was performed by SV40 viral transfection of the large T antigen into the primary muscle cells. Proliferation assays, growth curves, quantitative PCR, western blotting, mass spectrometry, and light microscopy were performed to characterize the MyoK9 cell line at different stages of growth and differentiation. The expression of muscle-related genes was determined to assess myogenic origin. Myok9 cells expressed dystrophin and other muscle-specific proteins during differentiation, as detected with mass spectrometry and western blotting. Using the Myok9 cell line, new therapies before moving to pre-clinical studies to enhance the number and speed of analyses and reduce the cost of early experimentation can be tested now. This cell line will be made available to the research community to further evaluate potential therapeutics.

中文翻译：

---

永生化的犬成肌细胞系：Myok9的创建和表征。

体外犬细胞系的可用性将减少狗对初级体外细胞培养的需求，并减少临床前研究的总成本。这组作者开发了一种永生化的犬肌肉细胞系，名为Myok9，来自正常犬的原代成肌细胞。永生化是通过将大T抗原通过SV40病毒转染到原代肌肉细胞中来实现的。进行增殖测定，生长曲线，定量PCR，蛋白质印迹，质谱和光学显微镜检查以表征MyoK9细胞系在不同生长和分化阶段的特征。确定肌肉相关基因的表达以评估成肌起源。Myok9细胞在分化过程中表达了肌营养不良蛋白和其他肌肉特异性蛋白质，这是通过质谱和蛋白质印迹法检测到的。使用Myok9细胞系，现在可以测试进入临床前研究的新疗法，以增加分析的数量和速度并降低早期实验的成本。该细胞系将提供给研究团体，以进一步评估潜在的治疗方法。