转化生长因子β1对骨髓干细胞分化类髓核细胞的影响  

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

作　　者：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 

机构地区：[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

出　　处：《中国组织工程研究与临床康复》2011年第36期6722-6726,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：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)~~

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(0,1,10,20μg/L)的HG-DMEM无血清培养液诱导3,7,14,21d;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:10μg/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14d时蛋白聚糖的表达均较3,7,21d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10μg/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。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.

关 键 词：转化生长因子Β1 骨髓间充质干细胞 髓核 分化 大鼠 

分 类 号：R394.2[医药卫生—医学遗传学]