兔骨髓基质干细胞诱导成骨的实验研究

Experimental study on induced bone formation of rabbit marrow stromal cells

目的:研究兔骨髓基质干细胞(BMSCs)的生长特点和在诱导条件下的成骨特性。方法:使用密度梯度法分离骨髓基质细胞并进行培养,传2代后运用成骨诱导剂地塞米松、β-甘油磷酸钠(β-Gp)、L-坏血酸(AA)进行诱导,测试骨髓基质细胞的生长变化和成骨分化。结果:形态学表明骨髓基质细胞呈集落生长,有成纤维细胞样外观,诱导后细胞生长较前明显减慢,逐渐变为立方形、锥形或梭形。汇合时细胞呈铺石状重叠生长,进而形成“钙结节”样表现。染色及电镜观察可见细胞诱导前后有明显区别,细胞增殖吸光度(OD值)及细胞碱性磷酸酶测试表明,随着时间的延长,细胞增殖变慢,但分化能力增强(P<0.05)。结论:骨髓基质细胞作为构建组织工程骨的种子细胞在地塞米松、β-甘油磷酸、L-坏血酸诱导下成骨具有可靠的重复性。

Objective:To investigate the growth features and osteogenesis characteristics of cultured rabbit marrow stromal cells(BMSCs) under bone induction conditions.Methods:BMSCs were isolated from rabbit by using density gradient separation method.Dex,β-Gp and AA were added to different passaged BMSCs,and proliferation and osteogenic differentiation of them were observed.Results:The morphological features of attached MSCs indicated colonies with the appearance of fibroblastic cells.The induced cells proliferation became slow,but differenation grew stronger with the time (P<0.05).Conclusion:The induced BMSCs with Dex、β-Gp and AA is a reliable tool for the bone tissue engineer.