Progenitor Cells inHuman Skeletal Muscle

这项提案的长期目标是确定人类肌肉中的单核细胞， 用于优化肌营养不良和肌病的细胞治疗。我们的最新发现 实验室已经表明，人骨骼肌中特定细胞群可能与一种 另一方面，调节肌肉前体细胞的增殖和分化。使用各种方法， 纯化，并与项目1密切合作，该项目将对肌肉祖细胞进行平行研究， 斑马鱼，我们将破碎人类肌肉前体细胞的几个子集。这些分离的部分将 在骨骼肌修复的体外和体内模型中使用三种不同的范例进行测试， 肌病：肌营养不良蛋白、dysferlin和肌管蛋白缺乏症。本申请的总体目标是， 更好地定义人类骨骼肌中的单核细胞及其相互作用， 可以鉴定出适合于治疗的高度增殖的肌原性细胞部分， 应用.这一目标将通过以下具体目标实现： 目标1。人胎儿和成人骨骼肌中的单个核细胞群，研究其细胞 表面相似性、体外增殖能力和肌生成潜力。 目标2.用mRNA表达芯片鉴定高增殖的人肌源性祖细胞 修复能力。 目标3。测定分级分离的人肌肉祖细胞修复突变的肌肉细胞的能力， 体外 目标4。确定不同的单核细胞群的恢复和肌源性潜力， 人体肌肉中。 希望通过与项目1密切协作，我们将能够了解 控制人类肌肉细胞增殖、生肌承诺和修复患病肌肉的能力。通过 通过这些努力，我们将确定人体肌肉中最有效的细胞群， 证明了它们在肌营养不良症和肌病的临床前小鼠模型中的有效性。如果 成功，这些发现可能会在基于细胞的治疗领域向前迈出重要一步， 肌肉萎缩症

The long-term objective of this proposal is to define mononuclear cells in human muscle that hold promise for optimization of cell-based therapy for muscular dystrophy and myopathy. Recent findings from our laboratory have indicated that specific cell populations in human skeletal muscle may interact with one another, regulating proliferation and differentiation of muscle precursor cells. Using various methods of purification and by working closely with Project 1, which will perform parallel studies on muscle progenitors in zebrafish, we will fractionate several subsets of human muscle precursor cells. These isolated fractions will be tested in both in vitro and in vivo models of skeletal muscle repair using three separate paradigms of myopathy: dystrophin, dysferlin and myotubularin deficiency. The overall goal of this application is that by better defining the mononuclear cells in human skeletal muscle and their interactions with one another, it will be possible to identify a highly proliferative, myogenic cell fraction that is amenable to therapeutic applications. This goal will be achieved via the following specific Aims: Aim 1. Fractionate mononuclear cell populations in human fetal and adult skeletal muscles, study their cell surface similarities, proliferative capacity and myogenic potential in vitro. Aim 2. Use mRNA expression arrays to characterize human myogenic progenitors with high proliferative and repair capacity. Aim 3. Assay the capacity of fractionated human muscle progenitor cells to repair mutant muscle cells in vitro. Aim 4. Determine the restorative and myogenic potential of distinct mononuclear cell populations within human muscle in vivo. The hope is that by working in close synergy with Project 1, we will be able to understand the signals that control human muscle cell proliferation, myogenic commitment and ability to repair diseased muscle. By pursuing each one of these efforts, we will define the most effective cell population(s) in human muscle and demonstrate their effectiveness in pre-clinical mouse models of muscular dystrophy and myopathy. If successful, these findings are likely to make a significant step forward in the field of cell-based therapy for muscular dystrophy.