论文部分内容阅读
[目的]观察成骨生长肽(osteogenic growth peptid,OGP)基因转染兔骨髓基质干细胞后的表达及表达产物对骨髓基质干细胞向成骨细胞分化的影响。[方法]构建重组真核表达载体pcDNA3.1-OGP,并在脂质体介导下,将其导入兔骨髓基质干细胞。通过G418筛选获得阳性克隆。用RT-PCR方法检测OGP基因在骨髓基质干细胞内的表达。I型胶原和碱性磷酸酶的检测观察转染pcDNA3.1-OGP后骨髓基质干细胞向成骨细胞分化情况。[结果]成功构建真核表达载体pcDNA3.1-OGP,用RT-PCR方法及碱性磷酸酶和Ⅰ型胶原检测证实OGP基因能在骨髓基质干细胞中表达并促进其向成骨细胞分化。[结论]经pcDNA3.1-OGP转染的兔骨髓基质干细胞不仅可以表达OGP,而且具有向成骨细胞系分化的特性。
[Objective] To observe the expression of osteoblastic growth factor (OPGP) gene transfected rabbit bone marrow stromal stem cells and the effect of its expression product on the differentiation of bone marrow stromal stem cells into osteoblasts. [Method] The recombinant eukaryotic expression vector pcDNA3.1-OGP was constructed and transfected into rabbit bone marrow stromal cells under liposome-mediated. Positive clones were obtained by G418 screening. The expression of OGP gene in bone marrow stromal cells was detected by RT-PCR. Type I collagen and alkaline phosphatase detection of pcDNA3.1-OGP transfected bone marrow stromal cells differentiate into osteoblasts. [Results] The eukaryotic expression vector pcDNA3.1-OGP was successfully constructed. The expression of OGP gene in bone marrow stromal cells and its differentiation into osteoblasts were confirmed by RT-PCR, alkaline phosphatase and type I collagen. [Conclusion] Rabbit mesenchymal stem cells transfected with pcDNA3.1-OGP not only express OGP but also differentiate into osteoblasts.