骨形成蛋白-2基因在人骨髓基质细胞中的表达及对其成骨分化的作用

BMP2 expression in human transfected mesenchymal stem cells and its effect on os teoblastic differentiation

目的利用构建的人骨形成蛋白-2(BMP2)真核表达载体pcDNA3/BMP2,检测其转染人骨髓基质细胞后的表达及对其成骨分化的影响。方法酶切鉴定构建的真核表达载体pcDNA3/BMP2,利用脂质体介导的转染技术,将所构建的载体导入骨髓基质细胞中,体外单层培养。分别于转染后48h和4周采用原位杂交、免疫组化和碱性磷酸酶、钙化学染色方法检测BMP2的基因蛋白表达以及对骨髓基质细胞成骨分化的影响。结果pcDNA3/BMP2酶切片段的大小与理论相符。转染后细胞能检测到BMP2基因和BMP2蛋白表达,并促进成骨转化。结论pcDNA3/BMP2转染骨髓基质干细胞中可获得短暂和长期表达,并加强骨髓基质细胞的成骨分化能力。

Objective To construct eukaryotic expression vector of human bon e morphogenetic protein 2, verify its expression in transfected human mesenchymal stem cells (hMSCs) and its effect on hMSCs differentiation. Method The BMP2 gene was cloned into a eukaryotic expression vector pcDNA3. The recombinant was tran sfected into hMSCs by liposome. Immunnohistochemistry and in situ hybridization methods were used to identify the expression of BMP2 mRNA and protein; ALP and V on Kossa stains were performed to identify the differentiated effect on the hMSC s. Result The pcDNA3/BMP2 fragments were the same as the theory has stated. BMP2 were expressed and synthesized both in 48h and 4w, and the osteoblastic differe ntiation was promoted. Conclusion Transfection of pcDNA3/BMP2 can provide transi ent and persistent expression in hMSCs, and promote the MSCs differentiation.