锌及碱性成纤维细胞生长因子单独或联合应用体外培养骨髓间充质干细胞生长及增殖

背景：如何简便有效的在体外大量扩增骨髓间充质干细胞，已成为神经科学研究热点。 目的：观察锌及碱性成纤维细胞生长因子单独或联合应用对骨髓间充质干细胞体外生长及增殖的影响，以寻求体外快速、大量增殖骨髓间充质干细胞的有效方法。 方法：体外培养大鼠骨髓间充质干细胞。实验分为对照组(不加任何影响因子)、锌组、碱性成纤维细胞生长因子组、锌+碱性成纤维细胞生长因子组，分别用特定浓度的锌及碱性成纤维细胞生长因子单独或联合应用。观察细胞生长曲线，用四唑盐比色法观察存活和增殖能力，流式细胞仪检测细胞周期，进行细胞表面抗原的鉴定。 结果与结论：细胞生长曲线、四唑盐比色法、流式细胞仪结果均显示，第3~5天各组增殖能力达高峰，其中锌+碱性成纤维细胞生长因子组增殖能力最强(P < 0.05)。第7天，对照组及单独应用碱性成纤维细胞生长因子和锌组细胞增殖力均有所下降，仅锌+碱性成纤维细胞生长因子组细胞仍保持较强的增殖(P < 0.05)。锌+碱性成纤维细胞生长因子组细胞中处于S+G2+M期的百分率最大，与其他3组相比差异有显著性意义(P < 0.01)。结果证实，锌和碱性成纤维因子均有促进体外培养的骨髓间充质干细胞增殖的作用，联合应用锌和碱性成纤维细胞生长因子促增殖作用最强，可以作为体外扩增培养骨髓基质细胞的最佳培养条件。

Growth and proliferation of bone marrow mesenchymal stem cells through in vitro culture of Zinc, basic fibroblast growth factor alone or their combination

BACKGROUND: It is the hot spot of neuroscience research addressing how to amplify a great quantity of bone marrow mesenchymal stem cells (BMSCs) in vitro by a convenient and effective way. OBJECTIVE: To study the effects of Zinc and basic fibroblast growth factor (bFGF) alone or in combination on the growth and proliferation of BMSCs in vitro, and to search an effective method of rapidly amplifying a great quantity of BMSCs. METHODS: Rat BMSCs were cultured in vitro. This study was composed of control group (without any influential factor), Zinc group, bFGF group, Zinc + bFGF groups, which was respectively treated with Zinc and bFGF alone or in combination. Cell growth curve was observed. Cell survival and proliferation were observed using MTT assay. Cell cycle was detected using flow cytometer, and cell surface antigen was identified. RESULTS AND CONCLUSION: Cell growth curve, MTT assay and flow cytometer results have shown that cell reproductive activity reached a peak at 3-5 days, and Zinc + bFGF group showed the strongest ability (P < 0.05). At day 7, cell reproductive activity was decreased in the control group, bFGF group and Zinc group, and only cells in the Zine + bFGF group exhibited a strong reproductive activity (P < 0.05). The percentage of cells in S+G2+M phase was greatest in the Zinc + bFGF group, and significant differences were found as compared with other three groups (P < 0.01). Results have verified that Zinc and bFGF can contribute to BMSC proliferation in vitro. Combination of Zinc and bFGF exhibits strongest effects on promoting proliferation, and can be an optimal culture condition for BMSCs in vitro.