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目的:观察重组人骨形成蛋白-7(rhBMP-7)对小鼠骨髓间充质干细胞(BMSCs)向成骨细胞分化的影响。方法:采用密度梯度离心法分离骨髓,体外培养扩增,获得小鼠骨髓间充质干细胞,贴壁培养至第3代,向培养液中加入rhBMP-7,培养5 d后进行碱性磷酸酶(ALP)染色,ALP活性与骨钙素(OC)含量检测,并用RT-PCR法分析成骨细胞标志基因骨钙素(OC)与Ⅰ型胶原(Col-Ⅰ)表达情况,Western blot法检测Ⅰ型胶原(Col-Ⅰ)蛋白的分泌量。结果:rhBMP-7诱导组的骨髓间充质干细胞的ALP染色强度、ALP活性及OC含量明显升高,OC与Col-Ⅰ的mRNA表达水平以及Col-Ⅰ的蛋白表达水平均明显提高。结论:rhBMP-7可促进体外培养的小鼠BMSCs向成骨细胞方向分化。
Objective: To observe the effect of recombinant human bone morphogenetic protein-7 (rhBMP-7) on the differentiation of mouse bone marrow mesenchymal stem cells (BMSCs) into osteoblasts. Methods: Bone marrow was isolated by density gradient centrifugation and cultured in vitro. Mice bone marrow mesenchymal stem cells were obtained and cultured to the third generation. RhBMP-7 was added to the culture medium and then incubated with alkaline phosphatase (ALP), osteocalcin (OC) and osteocalcin (OC) were measured. The expressions of osteocalcin (OC) and collagen type Ⅰ (Col-Ⅰ) were detected by RT- Type Ⅰ collagen (Col-Ⅰ) protein secretion. Results: The ALP activity, ALP activity and OC content of bone marrow mesenchymal stem cells induced by rhBMP-7 were significantly increased. The mRNA expression of OC and Col-Ⅰ and the protein expression of Col-Ⅰ were significantly increased. Conclusion: rhBMP-7 can promote the differentiation of mouse BMSCs into osteoblasts in vitro.