【摘要】 目的 探讨骨形态发生蛋白-7(BMP-7)的过表达载体对骨髓间充质干细胞(BMSCs)成骨分化的影响。方法 取10例健康志愿者的骨髓组织，培养原代BMSCs。采用流式细胞仪检测BMSCs的表面蛋白，对BMSCs进行鉴定。利用基因编辑技术，构建BMP-7的过表达载体，通过慢病毒转染BMSCs,将BMSCs分成转染组和未转染组。反转录聚合酶链反应(RT-PCR)检测BMP-7以及成骨分化标志物碱性磷酸酶(ALP)、Runt相关转录因子2(Runx2)、骨钙素(OCN)的表达。并对两组细胞进行茜素红和ALP染色。结果 BMSCs中CD90和CD106的表达为60.2%和58.3%;CD33和CD45的表达为3.4%和2.6%。慢病毒转染BMSCs的转染滴度为3×10~8 TU/ml。RT-PCR结果：BMP-7以及成骨分化标志物ALP、Runx2和OCN的mRNA的相对表达量转染组要明显高于未转染组(P<0.05)。茜素红染色结果：转染组肉眼可见明显红色染色区域，镜下可见大量钙结节点。ALP染色结果：转染组肉眼可见紫色染色区域，镜下可见ALP染色呈强阳性。结论 慢病毒转染BMP-7可以促进BMSCs成骨分化。更多还原

【Abstract】 Objective To investigate the effect of bone morphogenetic protein-7(BMP-7) gene on the osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods The bone marrow tissue of 10 healthy volunteers was taken and the primary BMSCs were cultured.The surface protein of BMSCs was detected by flow cytometry, and the BMSCs was identified.The over expression vector of BMP-7 was constructed by gene editing technique.BMSCs was transfected by slow virus, and BMSCs was divided into two groups: transfection group and non-transfection group.The expression of BMP-7 and alkaline phosphatase(ALP),Runt relative transfection factor 2(Runx2) and osteocalcin(OCN) were detected by reverse transcription-polymerase chain reaction(RT-PCR).The cells of two groups were stained with Alizarin red and alkaline phosphatase.Results The expressions of CD90 and CD106 in BMSCs were 60.2% and 58.3%;the expressions of CD33 and CD45 were 3.4% and 2.6%.The titer of lentivirus transfection was 3×10~8 TU/ml.The results of the RT-PCR:the mRNA relative expression level of BMP-7 and ALP,Runx2 and OCN mRNA of osteogenic differentiation markers in transfection group were significantly higher than those in non-transfection group(P<0.05).Alizarin red staining results: in the transfection group, obvious red area staining could be seen with naked eye, and a large number of calcium nodes could be seen under the microscope.ALP staining results: in the transfection group, purple area staining was visible with naked eyes, and ALP staining was strongly positive under microscope.Conclusions Lentivirus transfection of BMP-7 can promote the differentiation of BMSCs into osteoblasts.更多还原