碱性成纤维细胞生长因子对羊骨髓间充质干细胞体外增殖和分化的影响

目的：探讨碱性成纤维细胞生长因子（bFGF）对羊骨髓间充质干细胞（BMSCs）体外增殖与分化的影响。方法：体外分离培养的羊原代BMSCs分为实验组和对照组,实验组添加bFGF(2 μg•L^-1)于L-DMED培养液中,对照组使用L-DMED常规培养液。分别测定2组BMSCs生长曲线以及碱性磷酸酶（ALP）活性。应用诱导剂对2组细胞进行成骨、成脂肪诱导,分别在显微镜下观察其分化潜能。 结果：单个核细胞接种大约1周成纤维细胞样集落开始出现,2周后形成的集落数量增加,每个集落的体积变大。当第1代BMSCs细胞生长到汇合期时,实验组细胞数是对照组的2.5倍。于第16天实验组ALP活性比对照组明显降低（P＜0.05）。2组BMSCs经成骨诱导培养3周后,经Von Kossa染色,已经成骨的细胞内均可见钙盐沉积;已经成骨的细胞内油红-“O”染色显示,成脂肪诱导培养2周后,2组BMSCs均能分化形成脂肪细胞,2组每高倍视野下脂肪细胞计数差异无显著性（P>0.05）。 结论：bFGF能提高羊BMSCs集落形成数量及扩增效率,并抑制其ALP活性,同时保持了BMSCs向成骨细胞、脂肪细胞的分化潜能。

Effect of basic fibroblast growth factor on sheep bone marrow mesenchymal stem cells proliferation and differentiation in vitro

Objective To evaluate the effect of basic fibroblast growth factor(bFGF) on sheep bone marrow mesenchymal stem cells(BMSCs) proliferation and differentiation in vitro.Methods The sheep BMSCs were isolated and expanded in vitro and divided into experimental group and control group.bFGF(2 μg·L-1) was added into cell culture medium of experimental group.In control group,no bFGF was added.The morphological differences of BMSCs between two group were observed.Growth curves and alkaline phosphatase(ALP) activity were assessed.In order to observe the influence of bFGF on differentiation potential of BMSCs,osteogenic and adipogenic inductions were conducted on BMSCs in two groups.Results Fibroblast colonies appeared 1 week after seeding of the mononuclear cells.2 weeks later,more colonies formed and each colony became larger.The cell number was 2.5 times of the control when the first passage cells grew to confluence and the ALP activity in experimental group was lower than that in control group(P<0.05).Osteogenic and adipogenic inductions were confirmed by Von Kossa and oil red-"O" staining respectively.Conclusion bFGF can increase sheep BMSCs colony and proliferation efficiency and inhibit ALP activity while retain the potential of osteogenic and adipogenic differentiation in vitro.