摘要
目的探讨骨髓间充质干细胞经成骨细胞诱导不同时间后的细胞性质。方法成人骨髓分离单个核细胞(MNC),体外培养获得间充质干细胞(MSC),传至第4代后加入地塞米松(DEX)、抗坏血酸和β-甘油磷酸,诱导成骨分化。诱导7d和14d后的细胞与正常成人成骨细胞分别加入不同浓度骨形态发生蛋白-2(BMP-2)和DEX,48h后检测细胞增殖、碱性磷酸酶(ALP)活性和骨钙素(OCN)的含量。结果BMP-2和DEX对诱导7d后的细胞有明显的刺激增殖和ALP活性的作用;对诱导14d后的细胞,BMP-2和DEX都不能刺激细胞增殖和ALP活性,但OCN含量明显增加。正常成人成骨细胞对BMP-2和DEX作用的反应与诱导14d的成骨样细胞基本一致,但其OCN绝对含量明显高于MSC来源的成骨细胞。结论骨髓MSC向成骨诱导7d和14d后细胞分化程度不同。诱导时间在MSC的成骨分化中是一个非常关键的因素,但其机制和最佳诱导时间尚需进一步研究。诱导14d后的MSC的成骨活性低于正常成人成骨细胞。
Objective To compare the characteristics of MSC-derived osteoblast-like cells in different inductive time period. Methods Mononuclear cells (MNC) were isolated from adult bone marrow and cultured, mesenchymal stem cell (MSC) were obtained from the adherent cells. Dexamethasone( DEX), sodium-β-glycerophosphate and ascorbic acid were added into the culture of the fourth passage cells to induce osteogenic differentiation. After induction for 7 and 14 days MSC-derived ostebblast-like cells and normal osteoblast from adult tibia were treated with different concentrations of BMP-2 and DEX. Fourty-eight hours later, cell proliferation (MTI' method), ALP (alkaline phosphatase) activity and osteocalcin (OCN) content were measured. Results Proliferation and ALP activity significantly increased in the 7 d inductive osteoblast-like cells after 48 h administrated with BMP-2 and DEX. However, for the 14 d osteoblast-like cells, Neither proliferation nor ALP activity increased OCN content could be induced by BMP-2 and DEX. The reactions of normal osteoblast to the BMP-2 and DEX treatment were similar to those of 14 d osteoblast-like cells. The OCN level of normal oseoblast was much higher than those of MSC-derived osteoblast-like cells. Conclusion The results suggested that the characteristics of 7 and ld d MSC-derived osteoblast-like cells were different and the length of inductive period was a key factor for MSC osteogenic differentiation. Further study is needed to optimize the period and to explore the mechanism. The osteogenic potentiality of MSC-derived osteoblast-like cells was lower as compared to normal osteoblast.
出处
《基础医学与临床》
CSCD
北大核心
2005年第9期802-806,共5页
Basic and Clinical Medicine
基金
国家自然科学基金青年基金(30200134)
关键词
间充质干细胞
成骨细胞
诱导时间
mesenchymal stem cell
osteoblast
inductive interval