STEM CELL THERAPY FOR DISEASES OF BONE IN A MOUSE MODEL

干细胞治疗小鼠模型中的骨疾病

• 批准号: 6797851
• 负责人: CHRISTOPHER NIYIBIZI
• 金额: $ 35.18万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-27 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6797851
• 关键词: SDS polyacrylamide gel electrophoresis; bone marrow; cell differentiation; disease /disorder model; enzyme linked immunosorbent assay; fluorescent in situ hybridization; genetically modified animals; green fluorescent proteins; high performance liquid chromatography; histology; immunofluorescence technique; laboratory mouse; morphometry; musculoskeletal disorder therapy; musculoskeletal regeneration; osteoblasts; osteocalcin; osteogenesis imperfecta; polymerase chain reaction; stem cell transplantation; tissue /cell culture

The focus of the present proposal is to utilize a mouse model of osteogenesis imperfecta (oim) as a model system to evaluate the potential of the bone marrow derived mesenchymal stem cells (BMSCs) to engraft and participate in repair and regeneration of bone. The mouse has a natural occurring mutation that results in non-expression of proa2(I) chains leading to the accumulation of al(I) homotrimers in tissues. The mouse exhibits osteopenia, cortical thinning and easy fracturing and is an excellent model for evaluating the potential of BMSCs as targets for the treatment of genetic and non-genetic diseases of bone. Recent clinical trial by Horwitz et al. using whole marrow in children with a severe form of OI, demonstrated that BMSCs may offer treatment options for O1. Therefore, the hypotheses to be tested are: BMSCs from normal donor mice administered systemically or locally into syngeneic recipient mice will engraft in the bones of the recipient mice, synthesize authentic bone extracellular matrix and contribute to the structural integrity of the host bone. The following specific aims will be used to test these hypotheses: 1) Demonstrate that the cells infused into oim mice will engraft in bone and in fracture sites created in oim mice 2) Demonstrate that the cells which engraft in bone differentiate into osteoblasts and synthesize the authentic bone extracellular matrix and 3) Demonstrate that the cells that engraft in bone contribute to the structural integrity of bone. To accomplish the above aims, BMSCs will be established from femurs and tibiae of normal donor mice and either marked with retroviruses expressing LacZ or GFP genes to aid in cell tracking or unmarked prior to infusion in oim mice. The fate of the infused cells will be tracked by following expression of the marker genes in tissue and by fluorescent in situ hybridization (fish). Differentiation of the transplanted cells into osteoblasts in vivo will be determined by co-localization of osteocalcin and marker genes and also by in situ hybridization. Synthesis of authentic extraceltular matrix by the infused cells will be analyzed by the determination of the presence of type I collagen comprised of 1 and 2 heterotrimers. Structural integrity of the host bone, will be determined by histophotometry, cross-linking, and collagen content and bone mineral density. The proposed studies may lead to the development of better treatments for genetic and non-genetic diseases of bone based on BMSCs.

本研究的重点是利用成骨不全（oim）小鼠模型作为模型系统来评估骨髓间充质干细胞（BMSCs）移植和参与骨修复和再生的潜力。小鼠有一个自然发生的突变，导致proa2(I)链不表达，导致组织中al(I)三聚体的积累。小鼠表现出骨质减少、皮质变薄和易骨折，是评估骨髓间充质干细胞作为治疗遗传性和非遗传性骨疾病的潜在靶点的极好模型。最近由Horwitz等人进行的一项临床试验表明，骨髓间充质干细胞可以为严重的成骨不全症儿童提供治疗选择。因此，需要验证的假设是：来自正常供体的骨髓间充质干细胞