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目的体外建立小鼠血管内皮祖细胞(endothelial progenitor cells,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、7 d后RAW264.7细胞VEGFR-2和CXCR4 mRNA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、7 d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 of mouse endothelial progenitor cells (EPCs) and osteoclast precursor cell RAW264.7 cells in vitro. Methods EPCs and RAW264.7 cells co-cultured as experimental group, added the same amount of cell culture medium but did not join EPCs cells as a control group. Cell proliferation was detected by Cell Counting Kit-8 (CCK-8) kit and the growth curve of RAW264.7 cells during co-culture was detected by measuring the optical density. The co-culture of co-culture The expression of VEGFR-2 and CXCR4 mRNA in RAW264.7 cells were detected after 1, 3, 5 and 7 days. The activity of osteoclasts was detected by TRAP staining. Results Compared with RAW264.7 alone, the proliferation of RAW264.7 cells was accelerated by co - culture of EPCs cells. There was a significant difference (P <0.01) in optical density between the two groups after 3,5,7 days of co - culture. Co-culture promotes the differentiation of RAW264.7 cells into osteoclasts. Conclusion EPCs can promote the proliferation and differentiation of host osteoclast precursors in in vitro co-culture system.