干细胞因子与粒细胞集落刺激因子预处理对大鼠骨髓间质干细胞增殖分化的影响  

Effect of SCF and G-CSF pretreatment on the proliferation and the differentiation of bone mesenchymal stem cells

作　　者：鲍翠玉[1] 郭军[1] 马业新[2] 郑敏[1] 赵骥[1] 

机构地区：[1]咸宁学院心血管疾病研究所,湖北咸宁437100 [2]华中科技大学同济医学院附属同济医院心内科,湖北武汉430030

出　　处：《中国病理生理杂志》2007年第7期1363-1367,共5页Chinese Journal of Pathophysiology

基　　金：湖北省自然科学基金资助项目(No2005ABA085);湖北省教育厅资助项目(NoT200606)

摘　　要：目的:探讨干细胞因子(SCF)和粒细胞集落刺激因子(G-CSF)促进骨髓间质干细胞(MSCs)增殖及促进其向心肌细胞分化的作用。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(25mg/L)标记的P4MSCs与心肌细胞共培养。在共培养的第1d至第5d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高于对照组(P<0.01),单细胞因子组MSCs的心肌特异性蛋白MHC及TnT的阳性表达亦高于对照组(P<0.01)。结论:SCF和G-CSF对MSCs具有促增殖、促分化的作用。AIM ： To investigate the effect of pretreatment of stem cell factor （SCF） and granulocyte colony - stimulating factor （G -CSF） on the proliferation and the differentiation of mesenchymal stem cells （MSCs） into cardiomyogenic cells. METHODS ： The MSCs, isolated primarily from bone marrow, and purified by passage culture, were obtained from the adult rats of four groups： the rats were pretreated by 5 daily injections of SCF; the rats were pretreated with G-CSF; the rats were pretreated with SCF and G - CSF; the rats were treated without any intervention. The 4th passage of MSCs was labeled by DAPI and cellular cycle analysis was conducted by flow cytometry before co - culture. The neonatal rat cardiomyocytes cultured for 3 days were co - cultured with DAPI - MSCs. The percentage of the differentiation of MSCs into cardiomyogenic cells during the five co - culture days was analyzed. The morphologic changes of MSCs and the proteins expression of cardiac myosin heavy chain （MHC） and troponin T （TnT） were recorded respectively with digital microscope camera system and immunofluorescence technique. The percentage of the differentiation of MSCs into cardiomyogenic cells was also calculated. RESULTS： The percentage of MSCs in G0/G1 phase in SCF/G -CSF group was significantly lower than that in SCF group, G - CSF group and the control group. The percentage of MHC protein - positive MSCs in SCF/G - CSF group was markedly higher than that in SCF group, G - CSF group and the control group, and that in SCF group and G - CSF group was significantly higher than control group. The percentage of TnT protein - positive MSCs in SCF/G - CSF group, SCF group and G - CSF group was significantly higher than that in control group. CONCLUSION： SCF and G - CSF show the ability to stimulate the proliferation of MSCs and induce MSCs to differentiate into cardiomyocytes. The combination of using SCF and G - CSF is more effective than using only SCF or G - CSF.

关 键 词：间质干细胞 粒细胞集落刺激因子 心肌细胞 

分 类 号：R541.4[医药卫生—心血管疾病]