Abstract:
Skeletal muscle, which is predominantly constituted by multinucleated muscle fibers, plays a pivotal role in sustaining bodily movements and energy metabolism. Myoblasts, which serve as precursor cells for differentiation and fusion into muscle fibers, are of critical importance in the exploration of the functional genes associated with embryonic muscle development. However, the in vitro proliferation of primary myoblasts is inherently constrained. In this study, we achieved a significant breakthrough by successfully establishing a chicken myoblast cell line through the introduction of the exogenous chicken telomerase reverse transcriptase (chTERT) gene, followed by rigorous G418-mediated pressure screening. This newly developed cell line, which was designated as chTERT-myoblasts, closely resembled primary myoblasts in terms of morphology and exhibited remarkable stability in culture for at least 20 generations of population doublings without undergoing malignant transformation. In addition, we conducted an exhaustive analysis that encompassed cellular proliferation, differentiation, and transfection characteristics. Our findings revealed that the chTERT-myoblasts had the ability to proliferate, differentiate, and transfect after multiple rounds of population doublings. This achievement not only furnished a valuable source of homogeneous avian cell material for investigating embryonic muscle development, but also provided valuable insights and methodologies for establishing primary cell lines.
摘要:
骨骼肌,主要由多核肌纤维组成,在维持身体运动和能量代谢中起着关键作用。成肌细胞,作为分化和融合成肌纤维的前体细胞,在探索与胚胎肌肉发育相关的功能基因方面至关重要。然而,原代成肌细胞的体外增殖固有地受到限制。在这项研究中,通过导入外源鸡端粒酶逆转录酶(chTERT)基因成功建立鸡成肌细胞系,随后进行严格的G418介导的压力筛选。这种新开发的细胞系,被指定为chTERT成肌细胞,在形态上与原代成肌细胞非常相似,并且在至少20代种群倍增的培养中表现出明显的稳定性,而没有发生恶性转化。此外,我们进行了详尽的分析,包括细胞增殖,分化,和转染特性。我们的发现揭示了chTERT成肌细胞具有增殖的能力,区分,并在多轮人口倍增后进行转染。这一成就不仅为研究胚胎肌肉发育提供了有价值的均质禽类细胞材料来源,但也为建立原代细胞系提供了有价值的见解和方法。