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P物质在BMSCs来源的成骨细胞与内皮细胞体外联合培养中的作用
引用本文:李富航,靳宇飞,毕龙,裴国献.P物质在BMSCs来源的成骨细胞与内皮细胞体外联合培养中的作用[J].生物磁学,2014(19):3615-3618.
作者姓名:李富航  靳宇飞  毕龙  裴国献
作者单位:第四军医大学西京医院骨科,陕西西安710032
基金项目:国家自然科学基金项目(81272011)
摘    要:目的:观察P物质(substanceP,SP)在BMSCs来源的成骨细胞与内皮细胞体外联合培养中的作用,研究P物质作用于种子细胞的最适浓度指导组织工程骨修复骨缺损。方法:采用新生新西兰大白兔胎兔(雌雄不限)密度梯度离心法分离骨髓间充质干细胞行体外培养和连续传代,获得较纯的BMSCs。取生长状态良好的第3代BMSCs行成骨诱导培养及成血管内皮细胞诱导培养并鉴定。将诱导7d的两种细胞按2:1比例混合培养,待细胞传至2代加入不同浓度的sP作为实验组,以正常未加sP的细胞培养基为对照组。培养后1、3、5、7d采用CCK-8法测定细胞增殖并绘制生长曲线,观察细胞生长数量,测定碱性磷酸酶活性及观察细胞周期分布。结果:浓度范围从1×10^-12-1×10^-6mol/L的sP对联合共培养的成骨细胞增殖和活性都有促进作用,在浓度为1×10^-8mol/L对联合共培养的成骨细胞增殖和活性的作用功效最强。结论:在体外直接联合共培养的体系中,SP对新种子细胞促进效果明显,其在1×10^-8mol/L对联合共培养的成骨细胞增殖和活性作用最强。

关 键 词:P物质  BMSCs  成骨细胞  内皮细胞  共培养

Study of SP on Effect of Osteoblasts and Vascular Endothelial Cells from Rabbits BMSCs Co-cultured
LI Fu-hang,JIN Yu-fei,BI Long,PEI Guo-xian.Study of SP on Effect of Osteoblasts and Vascular Endothelial Cells from Rabbits BMSCs Co-cultured[J].Biomagnetism,2014(19):3615-3618.
Authors:LI Fu-hang  JIN Yu-fei  BI Long  PEI Guo-xian
Institution:(The Orthopedics Institute, Xijing Hospital, The fourth Military Medical University, Xi ' an, Shaanxi, 710032, China)
Abstract:Objective: To study the effect of SP (Substance P) of osteoblasts and vascular endothelial cells from rabbits BMSCs co-cultured in vitro. To find the optimal concentration of SP on the new seed ceils to guide bone defect repairing in tissue engineering of bone. Methods: The BMSCs were obtained from new-born rabbits by using gradient centrifuge method and cultured in vitro. The third generation of BMSCs were induced into osteoblasts and vascular endothelial cells. The osteoblasts and vascular endothelial cells, after being induced for 7 days in a ratio of 2 to 1, were directly co-cultured, the co-cultured osteoblasts and vascular endothelial cells in the second generation were used, and then SP with different concentrations was added (experimental group). While the second generation of co-cultured cells without SP was used as a control (control group). The function of SP was assessed by CCK-8 examination assay and the growth curve was drawn on 1,3,5,7 days, then counted the cells. Determined the ALP activity and examined the change of cell cycle distribution. Results: SP of 1 × 10^-12-1 × 10^-6 mol/L had enhancing effects on the proliferation and activity of co-culture osteoblasts, while the concentration of 1 × 10^-8 mol/L had the strongest enhancing effects. Conclusion: In direct co-culture system in vitro, the effect of SP-promoting on the new seed cells was significant. SP can improve activity of co-cultured osteoblasts.
Keywords:Substance P  BMSCs  Osteoblast  Endothelial cell  Co-cultured
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