CAJ | 학술논문

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.
배경:성골생장태구유명현적촉진성골세포증식、분화、성숙적작용.목적:관찰성골생장태기인전염토골수기질간세포후적표체급표체산물대골수기질간세포향성골세포분화적영향.방법:구건중조진핵표체재체pcDNA3.1-성골생장태,병재지질체개도하,장기도입토골수기질간세포.통과G418사선획득양성극륭.RT-PCR방법검측성골생장태기인재골수기질간세포내적표체,병관찰전염pcDNA3.1-성골생장태후골수기질간세포향성골세포분화적정황.결과여결론:실험성공구건료진핵표체재체pcDNA3.1-성골생장태,전염pcDNA3.1-성골생장태적골수기질간세포가견성골생장태mRNA적표체,동시간포안산적분비수평증가,감성린산매활성증고.증실경pcDNA3.1-성골생장태전염적토골수기질간세포불부가이표체성골생장태,이차구유향성골세포분화적특성.
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.