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背景:成骨生长肽具有明显的促进成骨细胞增殖、分化、成熟的作用。目的:观察成骨生长肽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-成骨生长肽,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。RT-PCR方法检测成骨生长肽基因在骨髓基质干细胞内的表达,并观察转染pcDNA3.1-成骨生长肽后骨髓基质干细胞向成骨细胞分化的情况。结果与结论:实验成功构建了真核表达载体pcDNA3.1-成骨生长肽,转染pcDNA3.1-成骨生长肽的骨髓基质干细胞可见成骨生长肽mRNA的表达,同时羟脯氨酸的分泌水平增加,碱性磷酸酶活性增高。证实经pcDNA3.1-成骨生长肽转染的兔骨髓基质干细胞不仅可以表达成骨生长肽,而且具有向成骨细胞分化的特性。
BACKGROUND: Osteogenic growth peptide has the obvious function of promoting the proliferation, differentiation and maturation of osteoblasts. OBJECTIVE: To observe the expression of osteogenic growth factor (rhGH) after transfection of rabbit bone marrow stromal cells and the effect of expressed products on the differentiation of bone marrow stromal cells into osteoblasts. Methods: The recombinant eukaryotic expression vector pcDNA3.1-osteogenic growth peptide was constructed and transfected into rabbit bone marrow stromal cells under liposome-mediated. Positive clones were obtained by G418 screening. The expression of osteogenic growth factor peptide in bone marrow stromal stem cells was detected by RT-PCR. The differentiation of bone marrow stromal stem cells into osteoblasts after transfection with pcDNA3.1-osteogenic growth peptide was observed. RESULTS AND CONCLUSION: The eukaryotic expression vector pcDNA3.1-osteogenic growth peptide was successfully constructed. The expression of osteogenic growth peptide mRNA was observed in bone marrow stromal cells transfected with pcDNA3.1-osteogenic growth peptide, and the hydroxyproline Increased secretion, alkaline phosphatase activity increased. Confirmed that pcDNA3.1-osteogenic growth peptide transfected rabbit bone marrow stromal stem cells can not only express osteogenic growth peptide, but also has the characteristics of differentiation into osteoblasts.