中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (50): 8042-8047.doi: 10.3969/j.issn.2095-4344.2014.50.002

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

大鼠骨髓间充质干细胞分离与培养:第3代细胞增殖快纯度高

曾智谋1,范忠诚2,张  寿2,刘亦恒2,吴多庆2,王琮仁2   

  1. 1成都医学院第一附属医院脊柱外科,四川省成都市  610500;2中南大学湘雅医学院附属海口医院骨科中心,海南省海口市  570208
  • 收稿日期:2014-11-02 出版日期:2014-12-03 发布日期:2014-12-03
  • 通讯作者: 张寿,教授,中南大学湘雅医学院附属海口医院骨科中心,海南省海口市 570208
  • 作者简介:曾智谋,男,1988年生,四川省资阳市人,汉族,2014年中南大学毕业,硕士,医师,主要从事脊柱外科方面的研究。
  • 基金资助:

    口市重点科技项目(2011-0135,2014-082);海口市人民医院院内课题(2014-YQY-07)

Isolation and culture of rat bone marrow mesenchymal stem cells: passage 3 cells exhibit rapid proliferation and high purity

Zeng Zhi-mou1, Fan Zhong-cheng2, Zhang Shou2, Liu Yi-heng2, Wu Duo-qing2, Wang Cong-ren2   

  1. 1Department of Spine Surgery, the First Affiliated Hospital of Chengdu Medical College, Chengdu 610500, Sichuan Province, China; 2Department of Orthopedic Surgery, Affiliated Haikou Hospital, Xiangya School of Medicine, Central South University, Haikou 570208, Hainan Province, China
  • Received:2014-11-02 Online:2014-12-03 Published:2014-12-03
  • Contact: Zhang Shou, Professor, Department of Orthopedic Surgery, Affiliated Haikou Hospital, Xiangya School of Medicine, Central South University, Haikou 570208, Hainan Province, China
  • About author:Zeng Zhi-mou, Master, Physician, Department of Spine Surgery, the First Affiliated Hospital of Chengdu Medical College, Chengdu 610500, Sichuan Province, China
  • Supported by:

    The Key Technology Project in Haikou City, No. 2011-0135, 2014-082; The Project of Haikou People’s Hospital, No. 2014-YQY-07

摘要:

背景:进行组织工程研究的首要环节是培养出大量生物活性好、增殖速度快、细胞纯度高的种子细胞。
目的:分析全骨髓贴壁法培养大鼠骨髓间充质干细胞的生物学特性。
方法:采用全骨髓贴壁法分离、培养大鼠骨髓间充质干细胞,绘制第3代细胞生长曲线,观察各代细胞形态学变化,流式细胞分析法测定第2代、第3代细胞表面标志物(CD29、CD90、CD34、CD45)的表达。
结果与结论:经全骨髓贴壁法培养3代以内骨髓间充质干细胞均具有典型成纤维样细胞形态而且贴壁生长;第2代骨髓间充质干细胞表型CD29、 CD90、CD34、CD45表达分别为91.5%,96.2%,1.3%,8.0%,第3代骨髓间充质干细胞分别为98.2%,98.1%,0.6%,2.0%;细胞生长曲线结果表明,以细胞浓度8×107 L-1接种,细胞增殖速度快,获得的细胞总数多。综合形态学及细胞鉴定结果,第3代大鼠骨髓间充质干细胞活性好、增殖速度快、细胞纯度高,适合作为组织工程研究的种子细胞。


中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程


全文链接:

关键词: 干细胞, 骨髓干细胞, 骨髓贴壁法, 细胞培养, 形态学, 生长曲线, 鉴定

Abstract:

BACKGROUND: The first step of tissue engineering research is to produce a large number of seed cells with good biological activity, fast proliferation and high purity.
OBJECTIVE: To explore the biological characteristics of rat bone marrow mesenchymal stem cells isolated and cultured by the whole bone marrow adherence method.
METHODS: Rat bone marrow mesenchymal stem cells were isolated and cultured, and then passage 3 cells were enrolled to draw a cell growth curve. Morphological changes of cells at different passages were observed. Cells in passage 2 and passage 3 were characterized by their surface marker expressions of CD29, CD90, CD34 and CD45.
RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells at passages 1-3 exhibit classic morphology of fibroblast-like cells and presented plastic-adherent property. Phenotypes of P2 generation were detailed as: expressions of CD29, CD90, CD34, CD45 were 91.5%, 96.2%, 1.3%, and 8.0%, respectively. Phenotypes of passage 3 cells were detailed as: expressions of CD29, CD90, CD34, CD45 were 98.2%, 98.1%, 0.6%, 2.0%, respectively. The cell growth curve indicated the cell proliferation reached the peak at an inoculation 
concentration of 8×107/L. Overall, bone marrow mesenchymal stem cells at passage 3 with good viability, rapid proliferation and high purity can be used as seed cells for tissue engineering research.


中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程


全文链接:

Key words: bone marrow, mesenchymal stem cells, cell culture techniques, cells, cultured

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