中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (6): 980-984.doi: 10.3969/j.issn.1673-8225.2011.06.008

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

不同来源人羊膜间充质干细胞的混合培养

王兆福1，焦红亮1，2，3，李建斌1，单  泓1，郑文迪4，谷晨熙4，杜  英5，关方霞6，杨  波2，3   

  1. 1河南省红十字血液中心，河南省郑州市  450014
    2郑州大学第一附属医院神经外科，河南省郑州市 450052
    3河南省高等学校临床医学重点学科开放实验室，河南省郑州市  450052
    4郑州大学临床医学系，河南省郑州市 450051
    5郑州大学基础医学院微生物与免疫学教研室，河南省郑州市 450052
    6郑州大学生物工程系，河南省郑州市  450052
  • 收稿日期:2010-11-20 修回日期:2010-12-20 出版日期:2011-02-05 发布日期:2011-02-05
  • 通讯作者: 关方霞，教授，郑州大学生物工程系，河南省郑州市 450001 guanfangxia@126.com 杨波，教授，博士生导师，郑州大学第一附属医院神经外科，河南省郑州市 450052 yangbo96@126.com
  • 作者简介:王兆福，男，1965年生，河南省南阳市人，汉族，1983年中山大学毕业，副教授，主要从事血液成分应用和外周血来源的干细胞研究。
  • 基金资助:

    郑州大学211工程三期重点学科建设项目；河南省医学科技创新人才工程项目(2005018)。

Mixed culture of human amnion-derived mesenchymal stem cells from different sources

Wang Zhao-fu1, Jiao Hong-liang1,2,3, Li Jian-bin1, Shan Hong1, Zheng Wen-di4, Gu Chen-xi4, Du Ying5, Guan Fang-xia6, Yang Bo2,3   

  1. 1Henan Provincial Red Cross Blood Center, Zhengzhou  450014, Henan Province, China
    2Department of Neurosurgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou  450052, Henan Province, China
    3Open and Key Laboratory of Clinical Medicine of Henan Colleges and Universities, Zhengzhou  450052, Henan Province, China
    4Department of Clinical Medicine, Zhengzhou University, Zhengzhou  450001, Henan Province, China
    5Department of Microbiology and Immunology, Basic Medicine School of Zhengzhou University, Zhengzhou  450052, Henan Province, China
    6Department of Biomedical Engineering, Zhengzhou University, Zhengzhou  450052, Henan Province, China
  • Received:2010-11-20 Revised:2010-12-20 Online:2011-02-05 Published:2011-02-05
  • Contact: Guan Fang-xia, Professor, Department of Biomedical Engineering, Zhengzhou University, Zhengzhou 450052, Henan Province, China guanfangxia@126.com Yang Bo, Professor, Doctoral supervisor, Department of Neurosurgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China; Open and Key Laboratory of Clinical Medicine of Henan Colleges and Universities, Zhengzhou 450052, Henan Province, China yangbo96@126.com
  • About author:Wang Zhao-fu, Associate professor, Henan Provincial Red Cross Blood Center, Zhengzhou University, Zhengzhou 450014, Henan Province, China
  • Supported by:

    the Key Subjects of Zhengzhou University*; Henan Innovation Talent Project of Medical Science, No. 2005018*

PDF

435

被引次数

4

摘要：

背景：人羊膜间充质干细胞可诱导分化为神经样细胞，但在培养过程中发现干细胞增殖的数量不足。
目的：为了获得足够数量的人羊膜间充质干细胞，观察人羊膜间充质干细胞混合培养的生长情况及经碱性成纤维细胞生长因子诱导人羊膜间充质干细胞向神经样细胞的分化情况。
方法：将两个不同胎盘来源的羊膜分离培养、混合，采用细胞分离和培养技术获取人羊膜间充质干细胞，分离人羊膜间充质干细胞后传代培养，加入碱性成纤维细胞生长因子诱导分化。当细胞传至第3代时，随机取培养的6份人羊膜间充质干细胞(半量)为A组，6份人羊膜间充质干细胞(半量)为B组，将A、B组各剩余的半量人羊膜间充质干细胞混合为C组。3组细胞浓度均为1.0×107 L-1。以活细胞计数和MTT比色法比较3组细胞扩增数量，以免疫组织化学法检测羊膜间充质干细胞神经胶质细胞标志物、神经元特异性烯醇化酶和巢蛋白的表达。
结果与结论：混合人羊膜间充质干细胞之间有互相促增殖的作用。人羊膜间充质干细胞具有较强的可朔性，经碱性成纤维细胞生长因子诱导的人羊膜间充质干细胞可表达神经胶质细胞标志物、神经元特异性烯醇化酶和巢蛋白。

关键词: 羊膜, 间充质干细胞, 诱导, 混合培养, 细胞增殖

Abstract:

BACKGROUND: Human amnion-derived mesenchymal stem cells (HAMs) can be induced to differentiate into neuron-like cells, but the number of stem cells in the training process is insufficient.
OBJECTIVE: Observed HAMs the growth of mixed culture, and induced HAMs to the nerve-like cells by basic fibroblast growth factor (bFGF) to obtain a sufficient number of HAMs.
METHODS: Amnions from two placentas were separated and cultured to obtain HAMs, which were induced differentiation by adding bFGF. When cells were at the 3rd passage, cultured 6 HAMs (half dose) were randomly taken for the A group, 6 HAMs (half dose) were taken for the B group, the remaining half of the amount of the A, B group HAMs mixed as C group. Cell densities of 3 groups were 1.0×107/L. The cells proliferations of 3 groups were compared by live cell count and MTT colorimetric. The expressions of GFAP, neuronspecific enolase (NSE) and nestin were detected by immunohistochemistry.
RESULTS AND CONCLUSION: Mixing HAMs have mutual effect on the proliferation. HAMs have a strong retrospective, which can express GFAP, NSE, and nestin induced by bFGF.

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