细胞因子对骨髓间充质干细胞增殖及其向心肌细胞分化的影响

The effect of cytokines on proliferation and differentiation of bone mesenchymal stem cells into cardiomyogenic cells

目的探讨干细胞因子(SCF)和粒细胞集落刺激因子(G-CSF)预处理对骨髓间充质干细胞(MSCs)增殖分化的作用。方法将SD大鼠随机分成对照组、SCF组、G-CSF组和SCF+G-CSF联合处理组。流式细胞仪检测第4代MSCs细胞周期。用DAPI(25 mg/m l)标记的P4MSCs与心肌细胞共培养。在共培养的第1天至第5天用免疫荧光技术分别检测心肌特异性肌节肌球蛋白重链(MHC)、肌钙蛋白T(TnT)的表达,统计向心肌样细胞分化的MSCs百分率。结果(1)SCF和G-CSF联合应用能明显诱导MSCs进入S期,单因子SCF组和G-CSF组与对照组相比,也明显的促进MSCs由G0/G1期进入S期(P<0.01)。(2)MSCs与心肌细胞共培养后,SCF和G-CSF联合组MSCs表达心肌特异性蛋白MHC及TnT的阳性百分率均显著增高,单细胞因子组MSCs的心肌特异性蛋白MHC及TnT的阳性表达亦较对照组高。结论SCF和G-CSF对MSCs具有促增殖、促分化的作用。

Objective To investigate the effect of pretreatment of stem cells factor （SCF） and granulocyte colony-stimulating factor （G-CSF） on proliferation and differentiation of mesenchymal stem cells （MSCs） into cardiomyngenic cells. Methods The SD rats were randomly divided into control, SCF, G-CSF and SCF ＋ G-CSF group. The 4th passage MSCs＇cellular cycle was valued by flowcytometry. The 4th passage of MSCs labeled by DAPI was co-cultured with cardiomyogenic cells. The proteins expression of cardiac myosin heavy chain （MHC） and troponin T （TnT） were detected with immunofluorescence technique from the first day to the fifth day to calculate the percentage of the differentiation of MSCs into cardiomyogenic cells. Resuits（1）The percentage of MSCs in G0/G1 phase of the SCF/G-CSF group was significantly lower than that of SCF ＋ G-CSF and control group （P 〈0. 01 ）. The percentage of MHC proteins？ positive MSCs of SCF/G-CSF group was significantly higher than that of SCF group, G-CSF group, and the control group, and that of SCF group and G-CSF group was significantly higher than that of the control group. （2）The positive percentage of MHC and TnT in SCF ＋ G-CSF group was significantly higher than that of SCF/G-CSF and control group, and that of SCF and G-CSF group was significantly higher than that of control group. Conclusions SCF and G-CSF could play the promotion role in proliferation and differentiation of MSCs.