Delivery of human muscle SP cells to dystrophic muscle

描述（由申请人提供）：肌肉SP细胞是一种原始细胞的异质群体，在体内表现出造血和肌源性分化潜力。这些细胞首次在小鼠中发现，有望优化肌营养不良症的细胞治疗。迄今为止，对人源性肌肉SP细胞及其假定的临床潜力知之甚少。该提案的目标是研究人胎儿和成人来源的肌肉SP细胞的分化潜力，并使用新生成的肌营养不良症小鼠模型优化其在临床前实验中的使用。这些目标将通过以下具体目标实现： 目标1.1。研究未分级分离的人肌源性SP和MP细胞在体外分化为肌源性或造血细胞的潜力。将从尸检胎儿和成人组织以及成人来源的废弃手术组织中获得人体样本。 目标1.2。基于细胞表面抗原CD 34、CD 90和CD 133的表达的人胎儿和成人肌肉SP和MP细胞的破碎。比较分离的和未分离的细胞在体外分化为肌源性或造血细胞的能力。 目标2.1。比较未分级和分级的肌源性SP和MP细胞在体内融合成营养不良肌纤维的能力。使用富含肌源性前体的胎儿和成人来源的人肌肉SP和MP细胞组分对NOD/RAG 1/null-DMD/mdx 5cv小鼠进行肌内注射（目的1.2）。 目标2.2。评估人胎儿和成人来源的肌肉SP或MP细胞与肌原性活性的临床潜力，在体内植入营养不良的骨骼肌。在非辐照NOD/RAG 1/null-DMD/mdx 5cv中进行人肌肉SP或MP细胞的动脉内和静脉内注射。 目标2.3。评估分级分离的人胎儿和成人来源的肌肉SP或MP细胞在体内分化为造血细胞的能力。将具有造血活性的未分级分离和分级分离的细胞（目的1.1和1.2）注射到致死性辐照的NOD/RAG 1/null/mdx 5cv或NOD/RAG 1/null-Pfp/null DMD/mdx 5cv小鼠的循环中。 这些研究将增强我们对人类肌肉SP细胞的基本知识，它们在胎儿和成人组织中的分化潜力，并将评估它们对肌营养不良症的治疗前景。

DESCRIPTION (provided by applicant): Muscle SP cells are a heterogeneous population of primitive cells that demonstrated hematopoietic and myogenic differentiation potential in vivo. First discovered in mice, these cells hold promise for optimization of cell-based therapy for muscular dystrophy. To date, little is known about human-derived muscle SP cells and their presumed clinical potential. The goals of this proposal are to study the differentiation potential of human fetal and adult-derived muscle SP cells, and to optimize their use in pre-clinical experiments using newly generated muscular dystrophy mouse models. These goals will be achieved via the following specific aims: Aim 1.1. Study the potential of unfractionated human muscle-derived SP and MP cells to differentiate into myogenic or hematopoietic cells in vitro. Human samples will be obtained from autopsy fetal and adult tissues and from adult-derived discarded surgical tissue. Aim 1.2. Fractionate human fetal and adult muscle SP and MP cells based on the expression of the cell surface antigens CD34, CD90 and CD133. Compare the ability of fractionated versus unfractionated cells to differentiate into myogenic or hematopoietic cells in vitro. Aim 2.1. Compare the ability of unfractionated and fractionated muscle-derived SP and MP cells to fuse into dystrophic myofibers in vivo. Perform intramuscular injections into NOD/RAG1/null-DMD/mdx5cv mice using the fetal and adult-derived human muscle SP and MP cell fractions enriched for myogenic precursors (Aim 1.2). Aim 2.2. Assess the clinical potential of human fetal and adult-derived muscle SP or MP cells with myogenic activity to engraft dystrophic skeletal muscles in vivo. Perform intra-arterial and intravenous injections of human muscle SP or MP cells in non-irradiated NOD/RAG1/null-DMD/mdx5cv. Aim 2.3. Assess the ability of fractionated human fetal and adult-derived muscle SP or MP cells to differentiate into hematopoietic cells in vivo. Inject unfractionated and fractionated cells with hematopoietic activity (Aims 1.1 and 1.2) into the circulation of lethally irradiated NOD/RAG1/null/mdx5cv or NOD/RAG1/null-Pfp/null DMD/mdx5cv mice. These studies will enhance our basic knowledge on human muscle SP cells, their differentiation potential in fetal and adult tissue, and will evaluate their therapeutic promise for muscular dystrophy.