TGF-β2和geneX对BrdU标记骨髓间充质干细胞增殖与成骨分化的作用

Effect of TGF- β2 and geneX on the Proliferation and Osteogenic Differentiation of Brd U- labeled Bone Mesenchymal Stem Cells

目的研究TGF-β2和gene X对Brd U标记的骨髓间充质干细胞增殖与成骨分化作用的影响.方法采用全骨髓贴壁培养法分离骨髓间充质干细胞,传代并鉴定,设实验组和空白对照组,实验组分4组:A组:Brd U标记后的BMSCs组;B组:Brd U标记后的BMSCs+TGF-β2组;C组:Brd U标记后的BMSCs+gene X组;D组:Brd U标记后的BMSCs+gene X+TGF-β2组.空白对照组为Brd U标记后的单纯BMSCs加入基础培养基避光培养.诱导培养后观察骨髓间充质干细胞生长形态、检测生长动力学(methyl thiazolyl tetrazolium,MTT)、碱性磷酸酶(ALP)和钙定量、进行细胞Von-Kossa法染色,观察成骨分化作用.结果 (1)4组BMSCs吸光度值(OD)分析比较显示:细胞组间差异有统计学意义(P<0.05);(2)各组BMSCs成骨诱导后碱性磷酸酶(ALP)和钙定量检测结果显示:细胞组间差异有统计学意义(P<0.05);(3)进行细胞Von-Kossa法染色,D组可见大量钙结节;C组可见较多钙结节;B组可见部分钙结节;A组可见少量钙结节;空白对照组未见钙结节.结论 (1)TGF-β2可促进骨髓间充质干细胞的增殖和诱导其向成骨细胞分化;(2)gene X可促进骨髓间充质干细胞向成骨细胞分化,且在细胞因子TGF-β2联合作用下成骨分化作用更明显.

Objective To study the effect of TGF- β2 and gene X on the proliferation and osteogenic differentiation of Brd U- labeled Bone Mesenchymal Stem Cells. Methods The whole bone adherent culture method was used to isolate, passage and identify bone mesenchymal stem cells. After grouped induction culture, the morphology, growth kinetics（ Methyl Thiazolyl Tetrazolium, MTT）, alkaline phosphatase（ ALP） and calcium quantification of BMSCs were observed and the osteogenic differentiation was observed by Von- Kossa staining.Results（ 1） The absorbance value（ OD） of BMSCs in four groups showed statistically significant differences among different cell groups（ P〈0.05）.（ 2） The alkaline phosphatase（ ALP） and calcium quantitative test results after osteogenesis induction showed statistically significant differences among different cell groups（ P〈0.05）.（ 3） Cell Von- Kossa staining showed that large amounts of calcium nodules in group D, more calciumnodules in group C, partial calcium nodules group B, a small amount of calcium nodules in group A, and no calcium nodules in control group. Conclusions（ 1） TGF- β2 can promote the proliferation of bone mesenchymal stem cells and induce them to differentiate into osteoblasts.（ 2） Gene X can promote bone mesenchymal stem cells to differentiate into bone cells, and osteogenic differentiation is more obvious in the cytokine TGF- β2 joint action.