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利用共培养法诱导人胎盘间充质干细胞向成骨细胞分化

Differentiation of human placenta derived mesenchymal stem cells into osteoblasts induced by co-culture method
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摘要 目的观察体外人足月胎盘间充质干细胞(h PMSCs)和成骨细胞共培养体系条件下成骨细胞对h PMSCs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h PMSCs进行鉴定。共培养组将成骨细胞接种于Transwell双层培养皿底层,h PMSCs接种于上层;对照组上层与底层均接种h PMSCs。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h PMSCs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h PMSCs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( h PMSCs) and osteoblasts in vitro and to study the influence of h PMSCs into osteoblasts in this system. Methods Mesenchymal stem cells( MSCs) were isolated and purified from human term placenta using collagenase digestion and identified by cell surface marker,cell growth curve,cell ultrastructure and osteogenesis. Osteoblasts were incubated in Transwell double-deck culture dish in the co-culture group. h PMSCs were incubated in the upper layer in high glucose DMEM medium with 10% FBS and identified by alkaline phosphatase staining. h PMSCs were incubated in both layers in the control group. Results Isolated placental cells can be identified as h PMSCs according to morphology,growth rate,cell surface 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-culture group was 21. 7% ± 5. 3%,the cells have the characteristics of osteoblasts,but in the control group negative. Conclusion h PMSCs have the same biological characteristics of MSCs from other sources and microenviroment provided by osteoblasts promotes the differentiation of h PMSCs into osteoblasts.
出处 《中华骨质疏松和骨矿盐疾病杂志》 2014年第4期325-331,共7页 Chinese Journal Of Osteoporosis And Bone Mineral Research
关键词 人胎盘间充质干细胞 成骨细胞 共培养 human placenta mensenchymal stem cells osteoblasts co-culture method
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