pcDNA3.1-osteogenic growth polypeptide eukaryotic expression vector in bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND: Osteogenic growth polypeptide (OGP) had clear effect on promoting osteoblast proliferation, differentiation and mature. OBJECTIVE: To explore the expression of OGP gene, which was transfected into rabbit bone marrow mesenchymal stem cells (BMSCs) and to evaluate the effects of OGP on differentiation of rabbit BMSCs. METHODS: pcDNA3.1-OGP was constructed using gene cloning and recombination techniques. Rabbit BMSCs were transfected with pcDNA3.1-OGP mediated by lipofectamine 2000. The transfection positive cell clones were selected with G418. The expression of OGP gene was detected using reverse transcription-polymerase chain reaction analysis on an mRNA level. Differentiation of pcDNA3.1-OGP transfected BMSCs into osteoblast lineage was observed. RESULTS AND CONCLUSION: The pcDNA3.1-OGP plasmid was constructed successful and OGP expression was detected in rabbit BMSCs. Hydroxyproline content was increased, and alkaline phosphatase activity was also increased. These indicate that pcDNA3.1-OGP transfected BMSCs expressed OGP, and could differentiate into osteoblast lineage.

Expression and biological effects of pcDNA_(3.1)-OGP gene transfection on rabbit bone marrow stromal cells in vitro / 中国矫形外科杂志

[Objective]To observe the expression of osteogenic growth peptide(OGP) gene,which had been transfected into rabbit bone marrow stromal cells(BMSC) by adenovirus,and detect the proliferation and differentiation of rabbit bone marrow stromal cells after gene transfection in vitro.[Methods]pcDNA3.1-OGP was constructed by using gene clone and recombined technique.With the help of lipofectamine 2000,BMSC were transfected with pcDNA3.1-OGP.The positive cell clones were selected with G418.The expression of OGP gene was observed by RT-PCR.Alkaline phosphatase(ALP) activity and type I collagen in pcDNA3.1-OGP transfected bone marrow stromal cells were measured to observe the differentiation from bone marrow stromal cells into osteoblast lineage.[Results]The pcDNA3.1-OGP vector was constructed successful.According to analysis of RT-PCR,alkaline phosphatase(ALP) and type I collagen assay showed that OGP gene could be expressed in BMSC,and could induce bone marrow stromal cells differentiation into osteoblast lineage.[Conclusion]The pcDNA3.1-OGP vector can be constructed successfully and its expression is confirmed in BMSC.The expression of OGP in bone marrow stromal cells can induce BMSC differentiation into osteoblast lineage.