胎盘来源间充质干细胞诱导分化为成骨细胞的实验研究

Experimental research on placenta-derived mesenchymal stem cells: Isolation,culture and differentiation towards osteoblast

目的：掌握体外分离、培养扩增胎盘间充质干细胞的方法，探讨其成骨细胞定向诱导分化的能力。方珐：用Ⅳ型胶原酶消化人胎盘组织，获得单个核细胞，接种于10％新生牛血清（FBS）低糖培养基中（DMEM），贴壁后常规换液、传代，流式细胞仪检测其表面标志，用β-甘油磷酸钠，维生素C，地塞米松联合诱导，使胎盘贴壁细胞分化为成骨细胞，诱导后10d检测碱性磷酸酶（ALP）活性，并行茜素红染色。结果：培养7～14d后有少量贴壁细胞生长，逐渐呈成纤维细胞状，表达CD29、CD44和CD105，不表达CD34、CD45、CD19；在成骨诱导10d后细胞ALP活性明显增强，茜素红染色可见钙盐沉积。结论：胎盘组织可能是新的间充质干细胞来源。

Objective： To explore the method for isolation, culture and expandition of placenta-derived mesenchymal stem cells（MSCs） in vitro. Methods： Mononuclear cells were harvested after placenta tissues were digested by eollagenaselV, and seeded in low-glucose DMEM containing 10%FBS, they were cultured and expanded in vitro, their markers were detected by flow cytometry; cells of 3 passages were induced to differentiate into osteoblast with vitamin C dexamethasone and β-glycerophoshate disodium salt-pentahydrate, induced cells were observed the change of alkaline phpsphatase activity and matrix mineralization. Results： B day 7-14 some adherent cells were observed and expanded same as spindle cells , the result showed that adherent cells were positive for CD29, CD44 and CD105, but negative for CD34, CD45, CD19; under the induced condition histological analysis of the cells revealed strong with alkaline phosphatase method and alizarin red confirmed that the cells had osteogenic potential. Conclusion： Placental mesenchymal stem cells could be regarded as an alternative source of MSCs.