中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (32): 5896-5900.doi: 10.3969/j.issn.1673-8225.2011.32.002

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

人骨髓间充质干细胞分离培养方法的改进

王承云1，石  磊2，夏  春2，郑欣鹏3，张  兵3，林飞太3，王少杰2   

  1. 1福建医科大学研究生院，福建省福州市  350108
    2厦门大学附属中山医院关节外科，福建省厦门市 361004
    3厦门大学医学院，福建省厦门市  361005
  • 收稿日期:2011-02-28 修回日期:2011-03-28 出版日期:2011-08-06 发布日期:2011-08-06
  • 通讯作者: 夏春，教授，博士，主任医师，硕士生导师，厦门大学附属中山医院关节外科，福建省厦门市 361000 Chunxia99@yahoo.com.cn
  • 作者简介:王承云★，男，1983年生，甘肃省兰州市人，汉族，福建医科大学在读硕士，主要从事骨与关节外科基础和临床研究。 hanson19830920@126.com
  • 基金资助:

    2009厦门科技基金资助课题子课题(3502Z20094008)，课题名称：股骨头坏死间充质干细胞成骨与凋亡过程中NF-kappaB调控caspase-3的研究。

Improvement of methods to isolate and culture human mesenchymal stem cells

Wang Cheng-yun1, Shi Lei2, Xia Chun2, Zheng Xin-peng2, Zhang Bing3, Lin Fei-tai3, Wang Shao-jie2   

  1. 1Fujian Medical University Postgraduate College, Fuzhou  350108, Fujian Province, China
    2Department of Joint Surgery, Zhongshan Hospital of Xiamen University, Xiamen  361004, Fujian Province, China
    3Xiamen University School of Medicine, Xiamen  361005, Fujian Province, China
  • Received:2011-02-28 Revised:2011-03-28 Online:2011-08-06 Published:2011-08-06
  • Contact: Xia Chun, Professor, Doctor, Chief physician, Master’s supervisor, Department of Joint Surgery, Zhongshan Hospital of Xiamen University, Xiamen 361004, Fujian Province, China Chunxia99@yahoo.com.cn
  • About author:Wang Cheng-yun★, Studying for master’s degree, Fujian Medical University Postgraduate College, Fuzhou 350108, Fujian Province, China hanson19830920@126.com
  • Supported by:

    Xiamen Technology Foundation Sub-project in 2009, No. 3502Z20094008*

PDF

468

被引次数

21

摘要：

背景：当前分离培养骨髓间充质干细胞方法的有多种，但在外科手术中进行获取的方法仍然较少，而外科分离所得细胞对于相应疾病的具有揭示原因和提供研究线索的价值。
目的：在骨科手术中以直接贴壁差速贴壁分离培养纯化与鉴定骨髓间充质干细胞。
方法：在骨科关节置换等大手术中吸取少量人骨髓血，采用直接贴壁法分离骨髓间充质干细胞，在贴壁后36~48 h行洗盘处理，并通过长期体外培养，特定培养基自身的筛选作用对细胞进行筛选，当细胞生长达到足量时进行鉴定。
结果与结论：采用直接贴壁法能够在长骨髓腔内混合血中分离培养骨髓间充质干细胞并形成集落，其形态学表现和表面分子经鉴定符合骨髓间充质干细胞特点，能够获得符合要求的人骨髓间充质干细胞，从而进行后续实验。

关键词: 人骨髓间充质干细胞, 细胞培养, 鉴定, 流式细胞术, 干细胞分离与培养

Abstract:

BACKGROUND: The methods to isolate mesenchymal stem cells (MSCs) can be classified to be several kinds. However, the method of acquisition in surgery is still less. The isolated MSCs from the patients who receive the replacement operation can offer some data which may reveal the causes or clues of some diseases.
OBJECTIVE: To isolate and culture MSCs with an improved technique of differential anchoring velocity and adherence, in order to identify MSCs.
METHODS: The bone marrow blood was collected in the femur marrow cavity duiring the hip joint replacement. MSCs were isolated by adherence method in the special medium and the mix medium was flushed 36-48 hours after the inoculation to isolate MSCs from the marrow mixture. In the long-time culture in vitro, the special medium was used to weed other cells out. The anchored cell was cultured and identified when the amount was large enough. 
RESULTS AND CONCLUSION: MSCs can be isolated from the bone marrow in long cavitas medullaris, and can be cultured into colonies by using adherence method. The morphological features and surface molecules are identified consistent with MSCs, and can obtain standard MSCs, so as to do the follow-up experiment.

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