目的体外建立小鼠血管内皮祖细胞(endothelial progenitorcells,EPCs)和破骨细胞前体细胞RAW264.7细胞株共培养体系。方法EPCs和RAW264.7细胞株共培养为实验组,加了等量细胞培养液但没有加入EPCs的细胞作为对照组。利用分离式培养小室进行细胞联合培养,Cell Counting Kit-8(CCK-8)试剂盒检测细胞的增殖,通过检测光密度值绘制共培养过程中RAW264.7细胞生长曲线;逆转录PCR检测共培养1、3、5、7d后RAW264.7细胞VEGFR-2和CXCR4mRNA的表达;TRAP染色检测破骨细胞活性。结果和EPCs细胞联合培养能加快RAW264.7细胞的增殖速率,和RAW264.7单独生长比较,共培养3、5、7d后,2组的光密度值差异有显著性(P〈0.01);共培养促进RAW264.7细胞分化为破骨细胞。结论在体外联合培养体系中,EPCs具有促进宿主破骨细胞前体细胞增殖与分化的作用。
Objective To establish a co-culture system containing mouse endothelial progenitor ceils (EPCs) and mouse osteoelast precursor RAW264. 7. Methods Co-cultured endothelial progenitor cells (EPCs) and RAW264.7 cells in Transwell chamber served as the experimental group, and the equivalent cells nutrient solution but without EPCs cells served as control group. Cell Counting Kit-8 was used to measure cell proliferation. Absorption values were detected to depicter cellular growth curve. Reverse transcript PCR was applied to detect VEGFR-2 and CXCR4 expression at mRNA level. TRAP staining was used to detect osteoclast activity. Results Co-culture system of mouse endothelial progenitor ceils (EPCs) and RAW264.7 cells in Transwell chamber facilitated RAW264.7 cells growth. In comparison with the growth of RAW264.7 cells in control group, the cell growth in co-culturing system was significantly increased after culture for 3, 5 and 7 d (P 〈0. 01 ). Co-culture also increased the expression of vWF and VEGFR-2 and the activity of osteoclast. Conclusion In our co-culture system of EPCs and RAW264.7 cells, EPCs facilitate the proliferation and differentiation of RAW264.7 cells.