中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (7): 985-990.doi: 10.3969/j.issn.2095-4344.2014.07.001

• 骨组织构建 bone tissue construction •    下一篇

骨髓贴壁细胞向破骨细胞的分化

陈  婧1,马  纪2,孙奋勇1   

  1. 1同济大学附属上海市第十人民医院检验科,上海市  200072
    2同济大学附属第十人民医院中心实验室,上海市  200072
  • 修回日期:2013-12-14 出版日期:2014-02-12 发布日期:2014-02-12
  • 通讯作者: 孙奋勇,博士,同济大学附属上海市第十人民医院检验科,上海市 200072
  • 作者简介:陈婧,女,1989年生,河南省新乡市人,汉族,同济大学在读硕士,主要从事骨分化及肿瘤方面的研究。
  • 基金资助:

    国家重大科学研究计划(2012CB966904)

Osteoclast differentiation of bone marrow adherent cells

Chen Jing1, Ma Ji2, Sun Fen-yong1   

  1. 1Department of Medical Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China
    2Department of Central Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China
  • Revised:2013-12-14 Online:2014-02-12 Published:2014-02-12
  • Contact: Sun Fen-yong, M.D., Department of Medical Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China
  • About author:Chen Jing, Studying for master’s degree, Department of Medical Laboratory, Tenth People’s Hospital, Tongji University, Shanghai 200072, China
  • Supported by:

    the National Key Scientific Research Project of China, No. 2012CB966904

摘要:

背景:一般认为采用贴壁分离法得到的小鼠骨髓中能够贴壁的细胞为间充质干细胞,并能分化成为成骨、成脂肪及成软骨细胞,其中贴壁的细胞是否有分化为破骨细胞的潜能尚不明确。
目的:检测贴壁分离法得到的小鼠骨髓中能贴壁的细胞是否能够分化为破骨细胞。
方法:采用贴壁法得到小鼠原代间充质干细胞,以及通过贴壁1-5 d后得到不同时间贴壁的骨髓细胞。原代间充质干细胞及不同时间贴壁的骨髓细胞分别用普通培养基,及含m-csf和RANKL培养基,培养9 d后进行碱性磷酸酶和抗酒石酸酸性磷酸酶染色。原代间充质干细胞传代后,第2代间充质干细胞分为4组,分别用普通培养基、含m-csf培养基、含RANKL培养基及含m-csf和RANKL的培养基培养,9 d后进行碱性磷酸酶及抗酒石酸酸性磷酸酶染色。
结果与结论:贴壁法得到较均一的间充质干细胞,原代及传代贴壁细胞的联合诱导组抗酒石酸酸性磷酸酶染色均为阳性,说明原代和传代的贴壁骨髓细胞中均有可以分化为破骨细胞的细胞。不同时间贴壁的细胞诱导后碱性磷酸酶及抗酒石酸酸性磷酸酶染色有差异,说明不同时间贴壁的细胞存在分化差异。



中国组织工程研究
杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程


全文链接:

关键词: 组织构建, 骨细胞, 骨髓间充质干细胞, 破骨细胞, 造血细胞, 贴壁分离法, 分离, 培养, 诱导, 分化

Abstract:

BACKGROUND: Generally considered bone marrow cells obtained by adherent method are mesenchymal stem cells, and they can differentiate into osteoblasts, adipocytes and chondrocytes.
OBJECTIVE: To explore the differentiation capacity of bone marrow adherent cells into osteoclasts.
METHODS: Primary mouse mesenchymal stem cells were obtained using adherent method, and bone marrow cells were obtained through adherence 1-5 days. Both of them were cultured with normal medium and inducing medium containing m-csf and RANKL. After 9 days, cells were stained by alkaline phosphatase and tartrate-resistant acid phosphatase. The passage 2 mesenchymal stem cells were divided into four groups, which were induced by mock, m-csf, RANKL and m-csf+RANKL, respectively. After 9 days of culture, the cells were stained by alkaline phosphatase and tartrate-resistant acid phosphatase.
RESULTS AND CONCLUSION: Primary culture of adhered cells was uniform. Alkaline phosphatase and tartrate-resistant acid phosphatase staining of primary and passaged bone marrow adherent cells induced with m-csf+RANKL were positive. This result showed that there were cells that could be induced into osteoclasts in the primary and passaged bone marrow adherent cells. Alkaline phosphatase and tartrate-resistant acid phosphatase  staining showed differences of adherent cells at different times after the induction, indicating that adherent cells at different times had different differentiation capacity.



中国组织工程研究
杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程


全文链接:

Key words: stem cells, mesenchymal stem cells, osteoclasts, cells, cultured

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