中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (27): 5023-5027.doi: 10.3969/j.issn.1673-8225.2010.27.019

• 干细胞因子及调控因子 stem cell factors and regulatory factors • 上一篇    下一篇

锌及碱性成纤维细胞生长因子单独或联合应用体外培养骨髓间充质干细胞的生长及增殖

任力杰，王  洋，韩漫夫   

  1. 深圳市第二人民医院神经内科 518035
  • 出版日期:2010-07-02 发布日期:2010-07-02
  • 作者简介:任力杰☆，男， 1972年生，山东省德州市人，汉族，2010南方医科大学毕业，博士，副主任医师，主要从事脑血管病、肌张力障碍及心理疾病研究。jesser2005@126.com
  • 基金资助:

    课题获广东省建设中医药强省科研课题(2009319)资助。课题题目：联合应用丹参酮ⅡA和人骨髓间充质干细胞移植治疗脑梗死的实验研究。

Growth and proliferation of bone marrow mesenchymal stem cells through in vitro culture of Zinc, basic fibroblast growth factor alone or their combination  

Ren Li-jie, Wang Yang, Han Man-fu   

  1. Department of Neurology, Shenzhen Second People’s Hospital, Shenzhen  518035, Guangdong Province, China
  • Online:2010-07-02 Published:2010-07-02
  • About author:Ren Li-jie☆, Doctor, Associate chief physician, Department of Neurology, Shenzhen Second People’s Hospital, Shenzhen 518035, Guangdong Province, China jesser2005@126.com
  • Supported by:

    the Scientific Research Foundation for Establishing Traditional Chinese Medicine Research of Guangdong Province, No. 2009319*

PDF

327

被引次数

16

摘要：

背景：如何简便有效的在体外大量扩增骨髓间充质干细胞，已成为神经科学研究热点。
目的：观察锌及碱性成纤维细胞生长因子单独或联合应用对骨髓间充质干细胞体外生长及增殖的影响，以寻求体外快速、大量增殖骨髓间充质干细胞的有效方法。
方法：体外培养大鼠骨髓间充质干细胞。实验分为对照组(不加任何影响因子)、锌组、碱性成纤维细胞生长因子组、锌+碱性成纤维细胞生长因子组，分别用特定浓度的锌及碱性成纤维细胞生长因子单独或联合应用。观察细胞生长曲线，用四唑盐比色法观察存活和增殖能力，流式细胞仪检测细胞周期，进行细胞表面抗原的鉴定。
结果与结论：细胞生长曲线、四唑盐比色法、流式细胞仪结果均显示，第3~5天各组增殖能力达高峰，其中锌+碱性成纤维细胞生长因子组增殖能力最强(P < 0.05)。第7天，对照组及单独应用碱性成纤维细胞生长因子和锌组细胞增殖力均有所下降，仅锌+碱性成纤维细胞生长因子组细胞仍保持较强的增殖(P < 0.05)。锌+碱性成纤维细胞生长因子组细胞中处于S+G2+M期的百分率最大，与其他3组相比差异有显著性意义(P < 0.01)。结果证实，锌和碱性成纤维因子均有促进体外培养的骨髓间充质干细胞增殖的作用，联合应用锌和碱性成纤维细胞生长因子促增殖作用最强，可以作为体外扩增培养骨髓基质细胞的最佳培养条件。

关键词: 锌, 骨髓间充质干细胞, 碱性成纤维细胞生长因子, 细胞增殖, 大鼠

Abstract:

BACKGROUND: It is the hot spot of neuroscience research addressing how to amplify a great quantity of bone marrow mesenchymal stem cells (BMSCs) in vitro by a convenient and effective way.
OBJECTIVE: To study the effects of Zinc and basic fibroblast growth factor (bFGF) alone or in combination on the growth and proliferation of BMSCs in vitro, and to search an effective method of rapidly amplifying a great quantity of BMSCs.
METHODS: Rat BMSCs were cultured in vitro. This study was composed of control group (without any influential factor), Zinc group, bFGF group, Zinc + bFGF groups, which was respectively treated with Zinc and bFGF alone or in combination. Cell growth curve was observed. Cell survival and proliferation were observed using MTT assay. Cell cycle was detected using flow cytometer, and cell surface antigen was identified.
RESULTS AND CONCLUSION: Cell growth curve, MTT assay and flow cytometer results have shown that cell reproductive activity reached a peak at 3-5 days, and Zinc + bFGF group showed the strongest ability (P < 0.05). At day 7, cell reproductive activity was decreased in the control group, bFGF group and Zinc group, and only cells in the Zine + bFGF group exhibited a strong reproductive activity (P < 0.05). The percentage of cells in S+G2+M phase was greatest in the Zinc + bFGF group, and significant differences were found as compared with other three groups (P < 0.01). Results have verified that Zinc and bFGF can contribute to BMSC proliferation in vitro. Combination of Zinc and bFGF exhibits strongest effects on promoting proliferation, and can be an optimal culture condition for BMSCs in vitro.

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