中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (19): 3042-3047.doi: 10.3969/j.issn.2095-4344.2014.19.015

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

原代人脐带间充质干细胞培养方法的研究

王  菲1，周  洪1，郭昱成1，苏晓霞1，李  昂1，赵小鹏2   

  1. 1西安交通大学口腔医院正畸科，陕西省西安市  710004
    2中国兵器工业第二零五研究所职工医院，陕西省西安市   710065
  • 修回日期:2014-03-08 出版日期:2014-05-07 发布日期:2014-05-07
  • 通讯作者: 周洪，教授，西安交通大学口腔医院正畸科，陕西省西安市710004
  • 作者简介:王菲，女，1975年生，山东省人，汉族，2013年西安交通大学毕业，博士，主治医师，主要从事骨组织工程的研究。
  • 基金资助:

    陕西省自然科学基础研究计划(2014JM4146)；陕西省“13115”科技创新工程计划项目(2008ZDKG-65)

Culture methods for primary human umbilical cord mesenchymal stem cells

Wang Fei1, Zhou Hong1, Guo Yu-cheng1, Su Xiao-xia1, Li Ang1, Zhao Xiao-peng2   

  1. 1Department of Orthodontics, Hospital of Dentistry, Xi’an Jiaotong University, Xi’an 710004, Shaanxi Province, China
    2The Hospital of No. 205 Research Institute of Weapons Industry in China, Xi’an 710065, Shaanxi Province, China
  • Revised:2014-03-08 Online:2014-05-07 Published:2014-05-07
  • Contact: Zhou Hong, Professor, Department of Orthodontics, Hospital of Dentistry, Xi’an Jiaotong University, Xi’an 710004, Shaanxi Province, China
  • About author:Wang Fei, M.D., Attending physician, Department of Orthodontics, Hospital of Dentistry, Xi’an Jiaotong University, Xi’an 710004, Shaanxi Province, China
  • Supported by:

    the Natural Science Foundation of Shaanxi Province, No. 2014JM4146; the “13115” Scientific Innovation Engineering Plan of Shaanxi Province, No. 2008ZDKG-65

PDF

895

被引次数

9

摘要：

背景：如何获得大量稳定活力好的细胞是人脐带间充质干细胞培养的难点。
目的：通过组织块法、酶消化法和酶解组织块法3种细胞培养方法比较，筛选出最佳的人脐带间充质干细胞的培养方式。
方法：分离新鲜脐带10根，分别采用组织块法、酶消化法、酶解组织块法3种方式培养人脐带间充质干细胞，比较3种方式原代人脐带间充质干细胞爬出所需时间、细胞培养成功率、绘制细胞增殖曲线，利用流式细胞仪检测细胞表面标志，并进行多项分化能力检测。
结果与结论：酶解组织块培养法原代人脐带间充质干细胞爬出所需时间与酶消化法无显著差异，但明显短于组织块法(P < 0.01)，原代人脐带间充质干细胞培养成功率显著高于其他两组，人脐带间充质干细胞增殖情况3组无显著性差异，酶解组织块培养法培养的第3代人脐带间充质干细胞其细胞表面标志及多项分化能力符合间充质干细胞的特性。酶解组织块培养法能缩短原代人脐带间充质干细胞爬出时间，显著提高原代人脐带间充质干细胞的成功率。


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, 脐带脐血干细胞, 间充质干细胞, 人脐带间充质干细胞, 原代细胞, 细胞培养, 组织块法, 酶消化法, 陕西省自然科学基金

Abstract:

BACKGROUND: How to get a lot of stable and dynamic human umbilical cord mesenchymal stem cells is still a difficulty.
OBJECTIVE: To investigate the best culture way of human umbilical cord mesenchymal stem cells by comparing tissue explant, enzyme digestion, enzymolysis methods.
METHODS: Ten fresh umbilical cords were isolated to human umbilical cord mesenchymal stem cells by using tissue explant, enzyme digestion, and enzymolysis methods, respectively. Hereafter, we compared cells harvested using three culture methods in terms of the time for the primary cells to creep, successful rates of cell-culturing, and curves of cell growth. Surface markers of cells were detected by flow cytometry and multiple differentiations of cells were detected.
RESULTS AND CONCLUSION: The time for the primary cells to creep out in the enzymolysis and enzyme digestion groups had no significant difference, but both of them were significantly shorter than that in the tissue explant group (P < 0.01). The successful rate of primary cell culture in the enzymolysis group was significantly higher than that in the other two groups. The cell proliferations of three groups had no significant difference. The cell surface markers and differentiation ability of the third generation of human umbilical cord mesenchymal stem cells cultured by enzymolysis method met the characteristics of mesenchymal stem cells. In conclusion, the enzymolysis method can shorten the time for primary human umbilical cord mesenchymal stem cells to creep out and increase the successful rates of primary human umbilical cord mesenchymal stem cells.


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: stem cells, mesenchymal stem cells, umbilical cord, cells, cultured

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