中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (36): 6722-6726.doi: 10.3969/j.issn.1673-8225.2011.36.017

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

转化生长因子β1对骨髓干细胞分化类髓核细胞的影响

呼  和1，2，韩成龙1，姜  超1，沈洪涛1，刘  杨1，闫  峰1，于长水1，永  梅3   

  1. 1哈尔滨医科大学附属第一医院骨科，黑龙江省哈尔滨市  150001
    2内蒙古民族大学医学院，内蒙古自治区通辽市  028000
    3内蒙古通辽市蒙中，内蒙古自治区通辽市  028000
  • 收稿日期:2011-03-02 修回日期:2011-05-23 出版日期:2011-09-03 发布日期:2011-09-03
  • 通讯作者: 韩成龙，博士后，副主任医师，硕士生导师，哈尔滨医科大学附属第一医院骨科，黑龙江省哈尔滨市 150001 Hanchenglong1973@126.com
  • 作者简介:呼和★，男，1979年生，内蒙古自治区通辽市人，蒙古族，哈尔滨医科大学在读硕士，内蒙民族大学医学院解剖教研室，主要从事脊柱外科及民族体育对人体身体素质的影响等研究。 huhe800@163.com
  • 基金资助:

    黑龙江省青年科学基金(QC2008C32)；黑龙江省博士后启动金、哈医大一院青年基金(2007097)。

Effect of transforming growth factor beta 1 on differentiation of bone marrow mesenchymal stem cells into nucleus pulposus-like cells

Hu He1,2, Han Cheng-long1, Jiang Chao1, Shen Hong-tao1, Liu Yang1, Yan Feng1, Yu Chang-shui1, Yong Mei3   

  1. 1Department of Orthopedics, First Affiliated Hospital of Harbin Medical University, Harbin 150001, Heilongjiang Province, China
    2School of Medicine, Inner Mongolia University, Tongliao  028000, Inner Mongolia Autonomous Region, China
    3Inner Mongolia High School, Tongliao  028000, Inner Mongolia Autonomous Region China
  • Received:2011-03-02 Revised:2011-05-23 Online:2011-09-03 Published:2011-09-03
  • Contact: Han Cheng-long, Doctor, Associate chief physicians, Master’s supervisor, Department of Orthopedics, First Affiliated Hospital of Harbin Medical University, Harbin 150001, Heilongjiang Province, China hanchenglong1973@126.com
  • About author:Hu He★, Studying for master’s degree, Department of Orthopedics, First Affiliated Hospital of Harbin Medical University, Harbin 150001, Heilongjiang Province, China; School of Medicine, Inner Mongolia University, Tongliao 028000, Inner Mongolia Autonomous Region, China huhe800@163.com
  • Supported by:

    the Youth Science Foundation of Heilongjiang Province, No. QC2008C32*; Heilongjiang Postdoctoral Startup Funding & Youth Funding of the First Affiliated Hospital of Harbin Medical University, No. 2007097*

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摘要：

背景：转化生长因子β1是骨髓间充质干细胞分化为类髓核细胞的首选生长因子，适当浓度能诱导骨髓间充质干细胞的增殖和分化。
目的：观察不同质量浓度转化生长因子β1对大鼠骨髓间充质干细胞体外向类髓核细胞分化的影响，优化骨髓间充质干细胞体外培养条件。
方法：取成年大鼠股骨骨髓，体外培养、纯化。取第3代成年大鼠骨髓间充质干细胞，实验组用加入不同质量浓度转化生长因子β1(0，1，10，20 μg/L)的HG-DMEM无血清培养液诱导3，7，14，21 d；对照组普通状态下用含体积分数为10%胎牛血清的HG-DMEM培养液自然分化。
结果与结论：10 μg/L转化生长因子β1诱导液诱导的骨髓间充质干细胞蛋白聚糖与Ⅱ型胶原蛋白水平明显高于其他实验组和对照组(P < 0.01)。各实验组14 d时蛋白聚糖的表达均较3，7，21 d时高(P < 0.01)。对照组各时间点蛋白聚糖Aggrecan及Ⅱ型胶原表达均呈阴性。提示10 μg/L的转化生长因子β1能提高大鼠骨髓间充质干细胞体外诱导分化类髓核的数量，从而使骨髓间充质干细胞发挥更高的治疗效率。

关键词: 转化生长因子&beta, 1, 骨髓间充质干细胞, 髓核, 分化, 大鼠

Abstract:

BACKGROUND: Transforming growth factor beta 1 (TGF-β1) is a first selected growth factor in differentiation of bone marrow mesenchymal stem cells (BMSCs) into nucleus pulposus (NP)-like cells. Suitable concentration can induce proliferation and differentiation of BMSCs.
OBJECTIVE: To observe the effects of different-concentration TGF-β1 on differentiation of BMSCs into NP-like cells, and to optimize culture conditions.
METHODS: BMSCs were separated and purified from adult rat femur bone marrow in vitro. The third-generated BMSCs were taken and induced with different concentrations of TGF-β1-contained in HG-DMEM culture medium in the experimental group, while in control group HG-DMEM medium containing 10% fetal bovine serum was used in general state. Immunohistochemical method was used to detect collagen type Ⅱ, and RT-PCR to detect the mRNA expression of collagen type Ⅱ and proteoglycan.
RESULTS AND CONCLUSION: The proteoglycans and collagen type Ⅱ of BMSCs was significantly higher than other experimental and control groups when TGF-β1 concentration was 10 μg/L (P < 0.01). The expression of proteoglycans at 14 days were higher than that at 3, 7, 21 days in the experimental groups (P < 0.01). The expression of proteoglycans and collagen type Ⅱ in the control group was negative. TGF-β1 can increase the number of differentiated BMSCs into NP-like cells at concentration of 10 μg/L, and make BMSCs play a greater treatment efficiency.

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