碱性成纤维细胞生长因子与猴骨髓间充质干细胞增殖及向神经元前体细胞分化:不同质量浓度对诱导剂隐丹参酮作用有差异吗?

Effect of basic fibroblast growth factor on proliferation and differentiation of monkey bone marrow mesenchymal stem cells into neuronal precursor cells:Does the concentration affect cryptotanshinone induction?

背景:碱性成纤维细胞生长因子属于活性单腱多肽,是一种有效的促有丝分裂因子。目的:探讨不同质量浓度碱性成纤维细胞生长因子对猴骨髓间充质干细胞增殖及成神经分化的影响。方法:密度梯度离心法体外分离培养猴骨髓间充质干细胞,传代后设立4组,对照组及低、中、高质量浓度组分别加入0,3,6,10μg/L碱性成纤维细胞生长因子,观察不同条件下猴骨髓间充质干细胞的增殖情况。取传至第5代骨髓间充质干细胞,用含20mg/L隐丹参酮的无血清L-DMEM培养基向神经方向诱导分化,免疫组化染色检测诱导后巢蛋白阳性的表达。结果与结论:与对照组比较,低、中、高质量浓度组骨髓间充质干细胞增殖速度明显加快(P<0.05),并且与培养液中碱性成纤维细胞生长因子的质量浓度呈正相关。隐丹参酮诱导0.5h后,低、中质量浓度组猴骨髓间充质干细胞均有一定程度的巢蛋白阳性表达;诱导1.5h后,巢蛋白阳性表达细胞数达峰值,低质量浓度组巢蛋白表达效果明显好于高质量浓度组(P<0.05)。提示碱性成纤维细胞生长因子可以促进猴骨髓间充质干细胞的体外增殖,并且在低质量浓度时能够增加隐丹参酮对骨髓间充质干细胞向神经元前体细胞分化的诱导率,在高质量浓度时则产生相反的抑制作用。

BACKGROUND： Basic fibroblast growth factor （bFGF） belongs to active peptide, which is an effective mitogenic factor. OBJECTIVE： To investigate the effect of bFGF on proliferation and differentiation of monkey bone marrow mesenchymal stem cells （BMMSCs） into neuronal precursor cells. METHODS： Monkey BMMSCs were in vitro cultured by density gradient centrifugation, and then divided into 4 groups after passaged, namely, control, bFGF with low, medium and high concentration groups. In the bFGF groups, 0, 3, 6, 10 μg/L bFGF were applied. The proliferation of BMMSCs in each group were observed. The 5th BMMSCs were cultured with serum free L-DMEM culture medium containing 20 mg/L cryptotanshinone to differentiated into neural-like cells. The expression of positive-nestin protein was detected by immunohistochemical method. RESULTS AND CONCLUSION： Compared with the control group, proliferation rate of BMMSCs in the bFGF groups were accelerated （P 0.05）, which showed a positive correlation to the concentration of bFGF. The positive-nestin protein could be found in the low and medium concentration groups at 0.5 hours after induction, and reached a peak at 1.5 hours, which increased obviously in the low concentration group than that of the high concentration group （P 0.05）. bFGF can promote BMMSCs proliferation in vitro, enhance inducing ratio of prophase neuron-like cells at lower concentration but inhibit differentiation at high level.