| 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].,2014,14(19):3615-3618 |
| 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 |
| Study of SP on Effect of Osteoblasts and Vascular Endothelial Cells fromRabbits BMSCs Co-cultured |
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| DOI： |
| 中文关键词: P物质 BMSCs 成骨细胞 内皮细胞 共培养 |
| 英文关键词: Substance P BMSCs Osteoblast Endothelial cell Co-cultured |
| 基金项目:国家自然科学基金项目（81272011） |
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| 中文摘要: |
| 目的：观察P 物质(substance P，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7 d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、7 d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。 |
| 英文摘要: |
| 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 cells 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. |
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