重组反转录病毒碱性成纤维细胞生长因子基因转染对人骨髓基质细胞成骨的影响

Effect of recombinant retrovirus pLXSN/basic fibroblast growth factor transfection on osteogenesis of human marrow stroma cells

背景:国内外有关成纤维细胞生长因子基因转染促血管和促肌肉生长的研究较多,而成纤维细胞生长因子基因促成骨的研究未见报道。目的:观察重组反转录病毒retroviruspLXSN/碱性成纤维细胞生长因子基因转染对人骨髓基质细胞成骨能力的影响。方法:从健康志愿者全骨髓中分离培养人骨髓基质细胞,体外扩增纯化后分为4组:①retroviruspLXSN/碱性成纤维细胞生长因子组:培养液中加入碱性成纤维细胞生长因子基因重组反转录病毒。②retroviruspLXSN组:培养液中加入反转录病毒空载体。③阳性对照组:培养液中添加地塞米松、抗坏血酸和β-甘油磷酸钠。④空白对照组:不给予特殊处理。结果与结论:经多次换液传代,人骨髓基质细胞呈均一梭形形态。处理后retroviruspLXSN/碱性成纤维细胞生长因子组与阳性对照组细胞形态逐渐趋于扁平,突起减少。免疫组织化学染色见retroviruspLXSN/碱性成纤维细胞生长因子组碱性成纤维细胞生长因子表达明显强于其他3组。retroviruspLXSN/碱性成纤维细胞生长因子和阳性对照组可引起细胞碱性磷酸酶活性增高和矿化结节及骨胶原形成。提示基因重组反转录病毒成纤维细胞生长因子转染对人骨髓基质细胞成骨能力具有促进作用。

BACKGROUND：In home and abroad have many studies on basic fibroblast growth factor（bFGF） transfection for improving the growth of muscle and vessel,but little studies on bFGF for improving the growth of osteogenesis.OBJECTIVE：To investigate the effect of recombinant retrovirus pLXSN/bFGF transfection on osteogenic potential of human marrow stroma cells（hMSCs）.METHODS：hMSCs isolated from health volunteers,cultured and purified in vitro were randomly divided into four groups.Retrovirus pLXSN/bFGF group with recombinant retrovirus pLXSN/bFGF was added into culture medium.Retrovirus pLXSN group with recombinant retrovirus pLXSN was added to take the place of retrovirus pLXSN/bFGF.Positive control group with dexamethasone,glycerophosphate and ascorbic were added into culture medium.Blank control group without any special treatment.RESULTS AND CONCLUSION：hMSCs were uniform spindle-shape fibroblast after several passages.After treatment,hMSCs in retrovirus pLXSN/bFGF and positive control groups gradually showed flat shape.Immunohistochemical staining showed that the level of bFGF expression in retrovirus pLXSN/bFGF group was higher than that of the other three groups.Mineralized nodules and bone collagen were formed,and alkaline phosphatase activity was improved in retrovirus pLXSN/bFGF and positive control groups.Recombinant of retrovirus pLXSN/bFGF gene transfection into hMSCs can promote the osteogenic potential of hMSCs.