›› 2016, Vol. 36 ›› Issue (5): 394-398.

• 基础研究 • 上一篇    下一篇

胎盘间充质干细胞牙向分化的体外研究

余永春,刘小峰,高平,袁林,杨征毅,孙洁   

  1. 广州医科大学附属第一医院
  • 收稿日期:2015-10-09 修回日期:2015-12-12 出版日期:2016-05-28 发布日期:2016-06-01
  • 通讯作者: 孙洁 E-mail:sjxj.558@163.com
  • 基金资助:
    骨形成蛋白信号通路在骨髓间充质干细胞牙向分化的作用和初步机制;广东省医学科学技术研究基金;胎盘间充质干细胞向成牙本质细胞分化的研究

A study on the odontogenic differentiation of placenta-derived mesenchymal stem cells in vitro

  • Received:2015-10-09 Revised:2015-12-12 Online:2016-05-28 Published:2016-06-01

摘要: [摘要] 目的探讨胎盘间充质干细胞牙向分化的可能性。方法双酶(胶原酶和胰蛋白酶)消化法获得SD大鼠胎盘间充质干细胞,免疫细胞化学鉴定其表型、成脂成骨诱导鉴定其多向分化能力。SD大鼠胎鼠牙胚细胞条件培养基诱导胎盘间充质干细胞14d,免疫细胞化学检测成牙本质细胞特异性蛋白-牙本质基质蛋白-1(Dentin matrix protein -1, DMP-1)和牙本质涎蛋白(Dentin sialoprotein, DSP),逆转录聚合酶链式反应(Reverse transcription polymerase chain reaction, RT-PCR)及凝胶电泳检测成牙本质细胞特异性基因-DMP-1和牙本质涎磷蛋白(Dentin sialophosphoprotein, DSPP)。结果胎盘间充质干细胞表达CD29、CD44和CD105,而不表达CD31、C34和CD45。成骨和成脂诱导后形成钙结节以及脂滴。牙向诱导后的胎盘间充质干细胞形态无明显改变,表达成牙本质细胞特异性相关蛋白-DMP-1、DSP和特异性基因-DMP-1和DSPP。结论胎盘间充质干细胞具有牙向分化的潜能。

关键词: 胎盘间充质干细胞, 成牙本质细胞, 牙胚细胞条件培养基, 分化

Abstract: Abstract: Objective To investigate the potential of odontogenic differentiation of placenta-derived mesenchymal stem cells (PMSCs). Methods PMSCs were obtained from SD rat placental tissues with collagenase and trypsin digestion. Immunocytochemistry was used to identify their phenotype. Osteogenic and adipogenic induction were used to identify their multilineage differentiation. SD fetal rat tooth germ cell-conditioned medium (TGC-CM) was used to induce PMSCs for 14 days. The specific proteins (DMP-1 and DSP) of odontoblast were detected by Immunocytochemistry. The specific genes (DMP-1and DSPP) of odontoblast were detected by RT-PCR and gel electrophoresis. Results PMSCs were positive for CD105, CD44 and CD29, negative for CD31, C34 and CD45. Calcium nodules and lipid droplets were formed respectively after osteogenic and adipogenic induction. After 14 days’ odontogenic induction, the morphology of PMSCs had no obvious change, and they expressed the specific proteins (DSP and DMP-1) and the specific genes (DMP-1 and DSPP), which were specifically expressed in odontoblast. Conclusion PMSCs have the potential to be induced to odontoblast-like cells.

Key words: Placenta-derived mesenchymal stem cells, Odontoblast, Tooth germ cell conditioned medium, Differentiation

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