碱性成纤维细胞生长因子及维生素C对骨髓间充质干细胞增殖的影响

Effects of Basic Fibroblast Growth Factor or Vitamin C on Proliferation of Bone Marrow Mesenchymal Stem Cells

目的研究碱性成纤维细胞生长因子(bFGF)及维生素C(Vc)两种影响因子单独或联合应用对骨髓间充质干细胞(BMSCs)体外生长及增殖的影响。方法体外培养大鼠骨髓间充质干细胞,分别用特定浓度的bFGF及Vc两种影响因子单独或联合应用,绘制细胞生长曲线,用四唑盐比色法(MTT)法、流式细胞仪及免疫组化染色法观察细胞的增殖及生长情况。结果细胞生长曲线、MTT法、流式细胞仪结果均显示第3～5天各组BMSCs增殖能力达高峰,其中Vc+bFGF组增殖能力最强(P<0.05)。第7天,对照组及单独应用bFGF和Vc组细胞增殖力均有所下降,仅Vc+bFGF组细胞仍保持较强的增殖能力(P<0.05)。结论bFGF和Vc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bFGF和Vc促增殖作用最强,可以作为体外扩增培养骨髓间充质干细胞的最佳培养条件。

Objective To study the effects of basic fibroblast growth factor （bFGF） or vitamin C used alone or in combination on proliferation of bone marrow mesenchymal stem cells （BMSCs） in vitro. Methods Rat BMSCs were cultured in vitro, and bFGF in definite concentration or vitamin C, alone or in combination, was added to the culture medium. Cell growth curves were obtained by cell counting, and cell proliferation and growth detected by MTr assay, flow cytometer and identification of cells origin. Results Cell growth curves, MTT assay and flow cytometer showed that proliferation reached the peak in all groups on days 3 - 5, and was the strongest in Vc ＋ bFGF group （ P 〈 0. 05 ）, and that on days 7, the proliferation decreased to some extent in groups control and alone - used bFGF or Vc, and remained relatively strong only in Vc ＋ bFGF group （ P 〈 0. 05）. Conclusion BFGF or Vc can promote the proliferation of BMSCs cultured in vitro, but the combined has the strongest effect which can be used as the best in vitro culture condition of BMSCs proliferation.