Delivery of human muscle SP cells to dystrophic muscle

描述(申请人提供)：肌肉SP细胞是一群异质性的原始细胞，在体内显示出造血和肌源性分化的潜力。这些细胞最先在小鼠身上发现，有望优化基于细胞的肌肉营养不良治疗。到目前为止，人们对人源性肌肉SP细胞及其推测的临床潜力知之甚少。这项建议的目标是研究人类胎儿和成人来源的肌肉SP细胞的分化潜力，并利用新产生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。研究未分离的人肌肉来源的SP和MP细胞在体外分化为肌源性细胞或造血细胞的潜能。人体样本将从尸检、胎儿和成人组织以及成人来源的废弃手术组织中获得。 目标1.2。根据细胞表面抗原CD34、CD90和CD133的表达，分离人胎儿和成人肌肉中的SP和MP细胞。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。比较分离的和分离的肌肉来源的SP和MP细胞在体内融合成营养不良的肌纤维的能力。对NOD/RAG1/Null-DMD/mdx5Cv小鼠进行肌肉注射，使用胎儿和成人来源的人类肌肉SP和MP细胞组分，富含成肌前体(AIM 1.2)。 目标2.2。评估具有肌源性活性的人胎儿和成人来源的肌肉SP或MP细胞在体内植入营养不良骨骼肌的临床潜力。在未照射的NOD/RAG1/Null-DMD/mdx5Cv中进行人肌肉SP或MP细胞的动脉内和静脉注射。 目标2.3。评估分离的人胎儿和成人肌肉SP或MP细胞在体内分化为造血细胞的能力。将分离的和分离的具有造血活性的细胞(AI1.1和1.2)注射到致死照射的NOD/RAG1/Null/mdx5Cv或NOD/RAG1/Null-PFP/Null DMD/mdx5Cv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，并将评估它们对肌肉营养不良的治疗前景。

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.