人骨形成蛋白-7基因在人骨髓间充质干细胞中的表达和对其分化表型的影响

目的：观察人骨形成蛋白-7(BMP-7)基因在人骨髓间充质干细胞(hMSC)中的表达及对其成骨表型的影响。方法：利用AdEasy腺病毒表达系统构建人BMP-7基因表达腺病毒AdB7V，体外感染hMSC后观察绿色荧光蛋白(GFP)表达及RT-PCR、免疫组化方法鉴定外源基因的表达，通过对感染细胞的碱性磷酸酶(ALP)染色和钙盐染色鉴定其成骨分化表型。结果：hMSC经AdB7V感染后72h可观察到GFP表达，RT-PCR、免疫细胞化学方法证实外源性BMP-7在hMSC中表达，感染细胞的ALP染色和钙盐染色均为阳性。结论：外源性的人BMP-7基因可在hMSC中表达并诱导其向成骨细胞分化。

Expression of human BMP-7 gene in mesenchymal stem cells and its effects on osteogenic phenotype

Objective: To investigate the expression of human BMP-7 gene in human mesenchymal stem cells(hMSCs) and its effects on osteogenic phenotype. Methods: The AdEasy system was used to generated the recombinant adenovirus AdB7V bearing human BMP-7 gene. The expression of exogenous genes in hMSCs after infection was determined by observing the expression of the green fluorescence protein(GFP) and detected by RT-PCR method and immunocytochemical staining. Identification of osteogenic phenotype was assayed by alkaline phosphatase(ALP) histochemistry and mineral deposition staining respectively. Results: GFP expression could be observed under fluorescent microscope 72h later after infection and expression of BMP-7 gene was confirmed by RT-PCR method and immunohistochemical staining. ALP and mineral deposition staining showed positive after infection. Conclusion: The results indicate that BMP-7 is expressed in hMSCs and is involved in inducing differentiation of hMSCs into osteoblast.