中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (27): 4964-4968.doi: 10.3969/j.issn.1673-8225.2011.27.006

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

骨髓间充质干细胞与髓核细胞的体外共培养

徐  远，俞  兴，徐  林   

  1. 北京中医药大学东直门医院骨科中心，北京市   100700
  • 收稿日期:2011-01-24 修回日期:2011-02-10 出版日期:2011-07-02 发布日期:2011-07-02
  • 通讯作者: 俞兴，博士，副主任医师，北京中医药大学东直门医院骨科中心，北京市 100700 并列通讯作者：徐林，博士，主任医师，博士生导师，博士后合作导师，北京中医药大学东直门医院骨科中心 100700
  • 作者简介:徐远★，男，1985年生，浙江省永康市人，汉族，2011年北京中医药大学毕业，硕士，主要从事脊柱应用基础研究。 xuyuan411@yahoo.cn

Co-culture of bone marrow mesenchymal stem cells and nucleus pulposus cells in vitro 

Xu Yuan, Yu Xing, Xu Lin   

  1. Orthopedic Center, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing  100700, China
  • Received:2011-01-24 Revised:2011-02-10 Online:2011-07-02 Published:2011-07-02
  • Contact: Yu Xing, Doctor, Associate chief physician, Orthopedic Center, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China Correspondence to : Xu Lin, Doctor, Chief physician, Doctoral supervisor, Orthopedic Center, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China
  • About author:Xu Yuan★, Master, Orthopedic Center, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, China xuyuan411@yahoo.cn

PDF

476

被引次数

15

摘要：

背景：细胞移植治疗椎间盘退行性变目前尚处在实验室研究阶段。
目的：通过SD大鼠骨髓间充质干细胞与退行性改变的髓核细胞体外共培养，探索骨髓间充质干细胞能否通过直接接触促进髓核细胞外基质的表达以及髓核细胞能否诱导骨髓间充质干细胞向类髓核细胞分化。
方法：取已自然退变的第6代髓核细胞与生长良好的第4代骨髓间充质干细胞按不同比例(75∶25，50∶50和25∶75)体外共培养7 d，单独髓核细胞(100∶0)与单独骨髓间充质干细胞(0∶100)分别作为阳性对照和阴性对照。
结果与结论：RT-PCR检测显示共培养组(75∶25和50∶50)蛋白多糖和Ⅱ型胶原蛋白条带均明显亮于单独髓核细胞培养组(100∶0)，吸光度值比较差异有显著性意义；免疫组织化学检测显示Ⅱ型胶原蛋白在共培养组(75∶25和50∶50)胞浆内的表达明显高于单独髓核细胞培养组(100∶0)。共培养组中可见骨髓间充质干细胞由长梭形向多角形、类圆形转变，胞浆内异染颗粒增多。提示通过体外直接接触共培养，骨髓间充质干细胞能逆转髓核细胞退变，而且髓核细胞能诱导骨髓间充质干细胞向类髓核细胞转换。

关键词: 髓核细胞, 骨髓间充质干细胞, 共培养, 椎间盘退行性变, 大鼠

Abstract:

BACKGROUND: Using stem cell transplantation in the treatment of disc degeneration disease is still limited to the laboratory study.
OBJECTIVE: To co-culture bone marrow mesenchymal stem cells (BMSCs) with the degenerative nucleus pulposus cells in vitro, and to explore whether BMSCs promote the expression of extracellular matrix of the nucleus pulposus cells and differentiate into nucleus pulposus-like cells in a directly contact microenvironment.
METHODS: The 6th generation of naturally degenerated nucleus pulposus cells were isolated and co-cultured with the 4th generation of BMSCs through direct contact in vitro, according to different proportions (75:25, 50:50, 75:25). The pure nucleus pulposus cells (100:0) and pure BMSCs (0:100) acted as the positive and negative controls, respectively.
RESULTS AND CONCLUSION: The results of reverse transcription-polymerase chain reaction of collagen II and aggrecan in the co-culture groups (75:25 and 50:50) showed brighter band than the positive control group (100:0), and there were significant differences of absorbance value. The results of immunohistochemical method of collagen II showed more expressions in the co-culture groups (75:25 and 50:50) than the positive control group (100:0). BMSCs turned into nucleus pulposus-like cells appearance and metachromatic granules increased in the co-culture group. Direct contact of MSCs and nucleus pulposus cell in vitro is essential for BMSCs reversing the nucleus pulposus cells degeneration and differentiating into nucleus pulposus-like cells.

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