大鼠骨髓基质细胞体外培养诱导成骨能力的研究  

Study on rat bone Marrow stroma cells cultured in vitro and osteogenic property

作　　者：高全文[1] 杨连甲[2] 梁立民[1] 柳春明[1] 

机构地区：[1]解放军总医院口腔颌面外科,北京市100853 [2]第四军医大学口腔医学院

出　　处：《中国美容医学》2005年第3期281-283,i003,共4页Chinese Journal of Aesthetic Medicine

摘　　要：目的:观察大鼠骨髓基质细胞在体外培养生长时的生物学特性,及诱导分化为成骨细胞的能力。方法:取成年大鼠2只,处死后分离、培养骨髓基质细胞,第8天时添加矿化液诱导培养,细胞经传代后绘制生长曲线。应用倒置相差显微镜、透射电镜观察其生长特性,并进行甲苯胺蓝染色,VonKossa染色,碱性磷酸酶染色。结果:培养的大鼠骨髓基质细胞呈成纤维细胞样生长,增殖活跃。经诱导后,细胞碱性磷酸酶活性高,连续培养30天,Von-Kassa染色显示有局灶状钙盐沉积。结论:MSCs易于分离培养,体外扩增速度快,在矿化条件下,骨髓基质细胞在体外即具有很强的成骨能力,为研究其作为骨组织工程种子细胞植入体内成骨奠定基础。Objective To observe the biological characterization and osteogenic property of rat bone Marrow stroma cells (MSCs ) cultured in vitro. Methods MSCs were obtained from the region of femur of rat, and were cultured in vitro. After 8 day, the adhesive cells were preserved to passage culture. MSCs were cultured in a conditional medium in subculture including:dexamethasone, vitamin C,and β-glycerophosphate.The cells proliferation capability and osteogenic property were examined by phase-contrast microscope,transmission electron microscope, histochemistry staining,Von Kossa straining and AKP straining. Results The adhesive MSCs were spindle shaped and had strong capability of proliferation.The cells occupied the whole bottom of cell culture bottle in about 5-6 days. Alkaline phosphate (AKP) activity of MSCs at passage 2 was increased significantly. When the cells were cultured up to 30 days,there was calcium deposition showed by von Kossa staining. Conclusion Rat MSCs have a strong activity of proliferation capability and osteogenic property. This study showed that rat MSCs can be used as seed cells in the bone tissue engineering.

关 键 词：诱导成骨能力 细胞体外培养 大鼠骨髓基质细胞 细胞碱性磷酸酶活性 组织工程种子细胞 碱性磷酸酶染色 透射电镜观察 甲苯胺蓝染色 生物学特性 相差显微镜 成骨细胞 诱导分化 成年大鼠 诱导培养 生长曲线 生长特性 连续培养 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学] R135.12[医药卫生—基础医学]