上海口腔医学 ›› 2015, Vol. 24 ›› Issue (3): 257-262.

• 基础研究 •    下一篇

干细胞表面标志物在牙髓干细胞中的表达

高丽1,江文欣1,牛晨光1,何智妍2,朱彩莲2,黄正蔚1   

  1. 1.上海交通大学医学院附属第九人民医院·口腔医学院 牙体牙髓病科,上海 200011;
    2.上海市口腔医学重点实验室,上海 200011
  • 收稿日期:2014-12-12 出版日期:2015-06-20 发布日期:2015-07-24
  • 通讯作者: 黄正蔚,Tel:021-23271699-5201,E-mail:huangzhengwei@shsmu.edu.cn
  • 作者简介:高丽(1986-),女,在读硕士研究生,E-mail:1127239423@sjtu.edu.cn
  • 基金资助:
    国家自然科学基金(81070826/81371143); 上海市启明星计划(12QH1401400)

Exploring the stem cell surface markers expressed in human dental pulp stem cells

GAO Li1,JIANG Wen-xin1,NIU Cheng-guang1,HE Zhi-yan2,ZHU Cai-lian2,HUANG Zheng-wei2   

  1. 1.Department of Endodontics, Shanghai Ninth People’s Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine. Shanghai 200011;
    2.Shanghai Key Laboratory of Stomatology. Shanghai 200011, China
  • Received:2014-12-12 Online:2015-06-20 Published:2015-07-24
  • Supported by:
    National Natural Science Foundation of China (81070826/81371143) and Shanghai Rising-Star Program (12QH1401400)

摘要: 目的:探讨磁珠分选后培养的人牙髓干细胞的表面标记抗原随培养代数增加的变化情况。方法:改良组织块法培养的人牙髓细胞,至第2代(P2)时,用磁珠方法分选出STRO-1阳性细胞,用流式细胞术分别检测P2、P3、P4、P5、P6、P7、P8代的干细胞表面标志物CD73、CD90、CD105、 CD166、STRO-1。取P3代细胞,分别进行成骨诱导和成脂诱导。21 d后,分别行茜素红染色和油红O染色,观察矿化物形成情况和脂滴形成情况,同时以未诱导细胞为对照。结果:STRO-1在牙髓干细胞表面随代数增加而下降,CD73、CD90、CD105、 CD166的表达比较稳定,茜素红染色可见矿化结节形成,油红O染色显示形成大量脂滴。结论:STRO-1在牙髓干细胞表面随代数增加而下降,其他干细胞标志物比较稳定。

关键词: 牙髓干细胞, 磁珠分选, 间充质干细胞, 表面标志物

Abstract: PURPOSE: To explore the stem cell surface markers expressed in human dental pulp stem cells which were selected and isolated by magnetic beads. METHODS: Human dental pulp cells (hDPCs) were separated and cultured from dental pulp of healthy third molars for orthodontic purpose. HDPSCs were isolated from cultured hDPCs by magnetic-activated cell sorting’s (MACS) indirect magnetic cell labeling and positive selection strategy with antibody STRO-1 in the 2nd generation. Then the stem cell surface markers (CD73, CD90, CD105, CD166 and STRO-1) were respectively detected in 3, 4, 5, 6, 7 and 8 generation of dental pulp stem cells. HDPSCs were induced to differentiation by adipogenic medium and osteogenic medium in the 3rd generation. Adipogenic differentiation was assessed by oil red O staining in day 21, and osteogenic differentiation was assessed by alizarin red staining in day 21. RESULTS: HDPSCs could differentiate into adipocyte and osteoblasts. Oil red O staining and alizarin red staining were positively expressed after induction of HDPSCs. STRO-1’s expression was decreased with the increase of generation. The expressions of CD73, CD90, CD105 and CD166 were relatively stable. CONCLUSIONS: The expression of STRO-1 is declined with the increase of generation, and the expressions of CD73, CD90, CD105 and CD166 are relatively stable with the changes of generation.

Key words: Human dental pulp stem cells, Magnetic-activated cell sorting, Mesenchymal stem cells, Surface markers

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