Progenitor Cells inHuman Skeletal Muscle

这项提案的长期目标是定义人类肌肉中有希望的单个核细胞 用于优化肌肉营养不良和肌病的基于细胞的治疗。来自我们的最新发现 实验室已经表明，人类骨骼肌中的特定细胞群可能与一种 另一个，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，我们将分离人类肌肉前体细胞的几个亚群。这些孤立的分数将 在体外和体内骨骼肌修复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、异铁蛋白和肌管蛋白缺乏症。这个应用程序的总体目标是通过 更好地定义人类骨骼肌中的单个核细胞及其相互作用，它将 有可能鉴定出一种高度增殖的、可用于治疗的肌源性细胞组分 申请。这一目标将通过以下具体目标实现： 目的1.人胎儿和成人骨骼肌中分离的单个核细胞群，研究其细胞 体外培养的细胞表面相似性、增殖能力和成肌潜能。 目的2.用基因表达谱芯片鉴定高增殖的人肌源性祖细胞 和维修能力。 目的3.检测分离的人肌祖细胞修复突变肌细胞的能力。 体外培养。 目的4.确定不同单个核细胞群体的修复和生肌潜能 活体中的人体肌肉。 希望通过与项目1密切协作，我们将能够理解 控制人类肌肉细胞的增殖、肌源性承诺和修复病变肌肉的能力。通过 在这些努力中，我们将定义人类肌肉中最有效的细胞群(S)和 在肌营养不良症和肌病的临床前小鼠模型中展示它们的有效性。如果 如果成功，这些发现可能会在基于细胞的治疗领域向前迈进一大步。 肌肉营养不良症。

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.