转化生长因子β1对骨髓干细胞分化类髓核细胞的影响

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。

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

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.