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山东大学学报(医学版)

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胎盘来源间充质干细胞诱导分化为成骨细胞的实验研究

孙念政,沈柏均   

  1. 山东大学齐鲁医院儿科, 山东 济南 250012
  • 收稿日期:2005-10-29 修回日期:1900-01-01 出版日期:2006-09-24 发布日期:2006-09-24
  • 通讯作者: 孙念政

SUN Nian-zheng,SHEN Bai-jun   

  1. Department of Pediatrics, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Received:2005-10-29 Revised:1900-01-01 Online:2006-09-24 Published:2006-09-24
  • Contact: SUN Nian-zheng

摘要: 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。方法: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10?d检测碱性磷酸酶(ALP)活性,并行茜素红染色。结果:培养7~14?d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10?d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。

Abstract: 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 collagenaseIV, 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.

Key words: Placenta, Mesenchymal stem cells, Differentiation, Osteoblasts

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