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首页> 外文期刊>Gene therapy >Retrovirally transduced bone marrow stromal cells isolated from a mouse model of human osteogenesis imperfecta (oim) persist in bone and retain the ability to form cartilage and bone after extended passaging.
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Retrovirally transduced bone marrow stromal cells isolated from a mouse model of human osteogenesis imperfecta (oim) persist in bone and retain the ability to form cartilage and bone after extended passaging.

机译：从人类成骨不全症（oim）小鼠模型中分离出的逆转录病毒转导的骨髓基质细胞在骨骼中持续存在，并在长时间传代后保留形成软骨和骨骼的能力。

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Bone marrow stromal cells isolated from a model of osteogenesis imperfecta (oim) mice, were transduced with a retrovirus (BAG) carrying the LacZ and neor genes after passage 21. The transduced cells retained the ability to express alkaline phosphatase activity in vitro when treated with recombinant human bone morphogenetic protein two (rhBMP-2), formed cartilage in vitro in aggregate cultures and formed bone in ceramic cubes after 6 weeks of implantation in nude mice. X-gal staining of ceramic cubes seeded with the transduced cells demonstrated the presence of LacZ-positive cells on the edges of bone and also in the lacunae of the newly formed bone 6 weeks after implantation. After infusion into femurs of oim mice, the transduced cells were detected in the marrow cavity and on the edges of the trabecular bone of the injected and contralateral femurs by X-gal staining and PCR analysis at 4, 10, 20, 30 and 40 days after injection. The LacZ gene was also detected in the lung and liver of the recipient mice at 4 and 10 days after injection but not at later time-periods. The present findings suggest that long-term cultured bone marrow stromal cells from osteogenesis imperfecta (OI) animals have the potential to traffic through the circulatory system, home to bone, form bone and continue to express exogenous genes. These findings open the possibility of using these cells as vehicles to deliver normal genes to bone as an alternative approach for the treatment of some forms of OI and certain other bone acquired and genetic diseases.
机译：从成骨不全症（oim）小鼠模型中分离出的骨髓基质细胞，在第21代传代后，携带带有LacZ和neor基因的逆转录病毒（BAG）进行转导。重组人骨形态发生蛋白2（rhBMP-2），在裸鼠体内植入6周后，在聚集培养物中体外形成软骨，并在陶瓷立方体中形成骨骼。用植入的转导细胞接种的陶瓷立方体的X-gal染色表明，植入后6周，骨边缘以及新形成的骨腔中存在LacZ阳性细胞。在输注oim小鼠的股骨后，通过X-gal染色和PCR分析在第4天，第10天，第20天，第30天和第40天时在注射的和对侧股骨的骨髓腔和小梁骨边缘检测到转导的细胞注射后。 LacZ基因还在注射后第4天和第10天在受体小鼠的肺和肝中检出，但在以后的时间中未检出。本研究结果表明，来自成骨不全症（OI）动物的长期培养的骨髓基质细胞有潜力通过循环系统运输，成为骨骼的家园，形成骨骼并继续表达外源基因。这些发现打开了使用这些细胞作为媒介物将正常基因传递给骨骼的可能性，作为治疗某些形式的OI和某些其他骨骼获得性遗传疾病的替代方法。

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