神经生长因子对大鼠骨髓基质细胞增殖和向成骨分化的影响
Effect of nerve growth factor on the proliferation and osteogenic differentiation of bone marrow stromal cells of rats
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摘要: 目的: 观察神经生长因子(NGF)对大鼠骨髓基质细胞(BMSC)增殖和向成骨分化的影响。方法: 体外培养大鼠BMSC,流式细胞术鉴定表面标志物;加成骨培养液培养至第3代,分为对照组(不加NGF)和实验组(加入100 ng/ml NGF),测定第1、3、5天第3代BMSC细胞增殖活性(MTT法)和碱性磷酸酶(ALP)活性,应用Von Kossa染色比较2组成骨性能。结果: CD90、CD105为强阳性表达,CD34、CD45为阴性。MTT法测得实验组第1、5天细胞的平均吸光度与对照组差异均无统计学意义(P＞0.05),而第3天实验组高于对照组(P＜0.05),实验组ALP表达明显高于对照组(P＜0.01);Von Kossa染色形成钙化结节也较对照组数量增多,面积增大。结论: NGF无明显促进大鼠BMSC增殖的作用,但可促进其向成骨细胞分化。Abstract: Objective: To investigate the effect of nerve growth factor(NGF)on the proliferation and osteogenic differentiation of bone marrow stromal cells(BMSC)of rats.Methods: The rat BMSC were cultured in vitro and the marker protein on the surface were identified by flow cytometry.The second generations of BMSC were cultured in osteogenetic medium.The third generation of BMSC were randomly divided into control group(no adding NGF)and experimental group(adding 100ng /ml NGF).The proliferation of the third generation of BMSC were detected by MTT assay, the activities of alkaline phosphatase (ALP)were observed and the osteogenic differatiation of the two groups were compared by von Kossa staining at days 1,3 and 5.Results: The postive CD90 and CD105 and negative CD34 and CD45 expression in cultured BMSC were detected by flow cytometry.Compared with the control group, the mean absorbance values of the experimental group at days 1 and 5 were not statistical different(P ＞ 0.05),but higher at day 3(P＜0.05),the activities of ALP was significantly higher(P＜0.01)and the number and area of the calcium nodules detected by von Kossa staining were larger.Conclusions: NGF can not promote the proliferation of BMSC,but can clearly promote the osteoblast differentiation
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