目的：观察体外人足月胎盘间充质干细胞（ hPMSCs ）和成骨细胞共培养体系条件下成骨细胞对hPMSCs分化的影响。方法采用胶原酶消化法从人足月胎盘中分离纯化间充质干细胞（ MSCs），检测细胞表面标志物、生长曲线、细胞超微结构及成骨能力并对hPMSCs进行鉴定。共培养组将成骨细胞接种于Tran-swell双层培养皿底层， hPMSCs接种于上层；对照组上层与底层均接种hPMSCs。对诱导后细胞进行碱性磷酸酶染色鉴定。结果胎盘分离细胞经形态、生长速度、细胞表面标志物（ CD44和CD29阳性表达为99％， CD34和CD106为1％），确定为胎盘间充质干细胞；头盖骨分离细胞经碱性磷酸酶染色确定为成骨细胞。采用Transwell共培养hPMSCs 和成骨细胞组碱性磷酸酶活性染色阳性率为（21.7±5.3）％，表现成骨细胞特性，对照组染色呈阴性。结论人足月胎盘含MSCs，与其他来源MSCs生物学特性相似，成骨细胞生长过程提供的微环境对hPMSCs分化为成骨细胞具有诱导促进作用。%Objective To establish the co-culture system between human placenta mensenchymal stem cells ( hPMSCs) and osteoblasts in vitro and to study the influence of hPMSCs into osteoblasts in this system .Methods Mes-enchymal stem cells ( MSCs) were isolated and purified from human term placenta using collagenase digestion and identi -fied by cell surface marker , cell growth curve , cell ultrastructure and osteogenesis .Osteoblasts were incubated in Tran-swell double-deck culture dish in the co-culture group .hPMSCs were incubated in the upper layer in high glucose DMEM medium with 10%FBS and identified by alkaline phosphatase staining .hPMSCs were incubated in both layers in the con-trol group.Results Isolated placental cells can be identified as hPMSCs according to morphology , growth rate, cell sur-face markers (CD44 and CD29 99%, CD34 and CD106 1%), ultrastructure, inducing ability , cells isolated from skull can determined as osteoblasts by alkaline phosphatase staining , positive rate of alkaline phosphatase staining in the co-cul-ture group was 21.7%±5.3%, the cells have the characteristics of osteoblasts , but in the control group negative .Con-clusion hPMSCs have the same biological characteristics of MSCs from other sources and microenviroment provided by osteoblasts promotes the differentiation of hPMSCs into osteoblasts .
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