人骨髓间充质干细胞体外成骨分化过程中成骨相关基因的动态表达

目的：系统研究人骨髓间充质干细胞（hBMSCs）体外成骨诱导分化过程中成骨相关基因的表达变化。方法：应用密度梯度离心法分离hBMSCs,取第2代细胞通过流式检测及多向诱导分化方法进行干细胞鉴定;应用RT-PCR法对hBMSCs在体外成骨诱导不同时间点的成骨相关基因表达进行检测。结果：第2代hBMSCs表达间充质干细胞表面标志CD44、CD90,具有成脂和成骨分化潜能。成骨相关基因在诱导早期部分表达,中期均有表达,基因表达大部分在14天达高峰,与矿化相关的基因表达在21天达高峰。结论：hBMSCs体外成骨诱导过程中成骨相关基因呈动态表达,其表达时序与成骨细胞生理发育基本相似。

Kinetic expression pattern of osteogenic genes during in vitro osteogenesis of human bone mesenchymal stem cells

Objective To investigate the osteogenic related genes expression during in vitro osteogenic differentiation of human bone mesenchymal stem cells（hBMSCs）.Methods hBMSCs were isolated by density gradient centrifugation.Surface markers and cell cycle were analyzed by flow cytometry.The multiple differentiation potential of hBMSCs were identified using in vitro osteogenic and adipogenic induction.The osteogenic related genes expression of hBMSCs at different induction time point were evaluated by semiquantitative RT-PCR analysis.Results The hBMSCs were derived from bone marrow and expressed CD44 and CD90,markers of mesenchymal stem cell.The second passage of hBMSCs showed adipogenic and osteogenic differentiation potential.During the osteogenic induction process,hBMSCs expressed part of osteogenic related genes in early stage,and all of them in middle stage.The peak expression of most of genes were on the 14th day,and the mineralization associated genes on the 21st day.Conclusions The osteogenic related gene expression of hBMSCs shows a dynamic progress during in vitro osteogenic induction,which is consistent with in vivo development of osteoblast.