人骨形态发生蛋白-2腺病毒表达载体转染人骨髓基质干细胞及对其增殖的影响

Effects of Transfection with an Adenoviral Vector Carrying the Human BMP-2 Gene on Human Bone Marrow Stromal Cells Proliferation in vitro

目的 :用人骨形态发生蛋白 - 2腺病毒表达载体 ( Ad- BMP- 2 )转染人骨髓基质干细胞( h BMSC) ,以探讨基因转染对 h BMSC增殖的影响。方法 :将 Ad- BMP- 2转染体外培养的成人骨髓基质干细胞 ,利用免疫组化、原位杂交染色方法检测细胞内骨形态发生蛋白 - 2 ( BMP- 2 )的表达 ,蛋白印迹法检测转染后细胞培养液中 BMP- 2分泌蛋白表达。然后通过流式细胞仪分析其对细胞增殖的影响。结果 :转染后 ,h BMP- 2基因在 m RNA水平和蛋白水平均有表达。蛋白印迹法检测到培养液中有 BMP- 2蛋白阳性表达。转染 h BMP- 2基因后 ,细胞经流式细胞仪分析 ,S期细胞比例增多 ,说明细胞 DNA的合成增加。结论 :Ad- BMP- 2可高效转染 h BMSC。

Objective:The Adenoviral vector carrying hBMP 2 was transfected into human bone marrow stromal cells (hBMSCs) in vitro to study the effects of the transfection on these cells proliferation. Methods:The expression of hBMP 2 in these cells after infection was determined by in situ hybridization and immunohistochemical analysis. BMP 2 productions were confirmed by western blot analysis of the supernatant collected from the cells. The changes in the cells proliferation were observed by flowcytometry. Results:The expression of hBMP 2 was confirmed in levels of mRNA and protein. The presence of BMP 2 was detected in the supernatant of infected cells. Cellular proportion in S period increased after infection, which indicates the increase of synthesis and propagation of cell DNA. Conclusion: Ad BMP 2 could infect hBMSC with high transduction efficiency and promote cells proliferation.