MUSCLE SATELLITE CELL--REACTION TO DISEASE AND TRAUMA

成肌细胞移植已被认为是一种基因互补的方法 用于治疗原发肌病并补充可能缺少的蛋白质 体循环。该系统还提供了一个独特的机会 评估肌肉发育和再生。建议研究一下 利用胎儿宿主和稳定的成肌细胞移植系统的细胞生物学 有标记的成肌细胞。胎儿宿主的使用将克服这些问题 成肌细胞转移到更成熟的宿主中所固有的问题(即 供体成肌细胞的传递、分散和整合为显著的 肌纤维数量和免疫排斥问题)，如 免疫功能低下胎儿发育中的小肌肉注射 东道主将进行广泛的从头肌管形成，并将有 组织不良的结缔组织。这一百万美元应该会导致 大量广泛分散的马赛克肌管的生产 (具有不同频率的可识别的供体成肌细胞)，没有 求助于诱导成肌细胞所需的肌肉创伤 整合到肌肉中，包含其全部的肌纤维。 这将允许评估注射的成肌细胞的命运， 确定它们的一些后代是否保留为肌卫星 细胞，能够参与肌纤维的生长和再生，并 供体来源的肌核长期存活和 继续生产独特的结构蛋白产品。这个问题是 成肌细胞对特定细胞谱系的承诺随后将通过 从胚胎肌肉中转移标记稳定的成肌细胞 肌卫星细胞从成人肌肉进入发育中的胎儿肌肉。我们会 还提到了使用成肌细胞移植作为一种方法的潜在好处 基因互补将dystrophin替换为 MDX胎鼠肌营养不良蛋白缺陷性肌纤维。供体频率 肌营养不良蛋白阳性的mdx鼠马赛克肌纤维内的肌核 以及抗肌营养不良蛋白的替代在改变 将评估肌营养不良蛋白缺乏症肌肉的变性-再生。 将通过免疫组织化学和光学显微镜进行评估。 和扫描共聚焦显微镜，以及免疫印迹抗体 抗肌营养不良蛋白和抗肌球蛋白抗体或诱导。结果是 将提供重要的新信息 成肌细胞转移系统将为未来的使用开辟道路 该系统用于治疗目的，并作为评估许多 肌肉发育的细胞生物学方面。

Myoblast transfer has been suggested as a method of gene complementation for primary myopathics and to supplement proteins that may be absent in the systemic circulation. This system also offers a unique opportunity to evaluate muscle development and regeneration. It is proposed to study the cell biology of the myoblast transfer system using fetal hosts and stably marked myoblasts. The use of fetal hosts will overcome the problems inherent in myoblast transfer into more mature hosts (i.e., problems of delivery, dispersal and integration of donor myoblasts into a significant number of myofibers and problems of immune rejection), as at the time of injection the small developing muscles of the immune-incompetent fetal hosts will be undergoing extensive de novo myotube formation and will have poorly organized connective tissue. This millieu should result in the production of a significant number of widely dispersed mosaic myotubes (with different frequencies of identifiable donor myoblast), without resorting to the muscle traumatization required to induce myoblast incorporation into muscles containing their full complement of myofibers. This will permit evaluation of the fate of the injected myoblast, determining whether some of their progeny are maintained as myosatellite cell, capable of participating in myofiber growth and regeneration and of the ability of donor-derived myonuclei to survive long-term and to continue to produce a unique structural protein product. The question of myoblast commitment to specific cell lineages will then be addressed by transferring stably marked myoblasts from embryonic muscle and myosatellite cells from adult muscle into developing fetal muscle. We will also address -the potential benefit of using myoblast transfer as a method of gene complementation for the replacement of dystrophin into the dystrophin-deficient myofibers of fetal mdx mice. The frequency of donor to host myonuclei in dystrophin-positive mosaic myofibers in the mdx mice and the effectiveness of dystrophin replacement on modifying the degeneration-regeneration of dystrophin deficient muscle will be assessed. Evaluations will be made with immunohistochemistry, and light microscopy and scanning confocal microscopy, and with Western blotting with antibody against dystrophin and antibodies against myosin or induction. The results of the proposed experiments will provide significant new information about the myoblast transfer system that will be open the way for future use of this system for therapeutic purposes and as a tool for evaluating many aspects of the cell biology of muscle development.