人骨髓成骨细胞体外培养  

In Vitro Culture of Human Osteoblasts from Bone Marrow

作　　者：黄柏英[1] 祝斌[1] 罗学滨[1] 吴尚辉[1] 祝和成[1] 

机构地区：[1]中南大学细胞中心,湖南长沙410078

出　　处：《实用预防医学》2004年第5期932-934,共3页Practical Preventive Medicine

摘　　要：目的　研究人骨髓基质干细胞体外培养及其成骨特性 ,为骨组织工程学研究提供成骨种子细胞。　方法　以人髂骨区骨髓为材料进行体外人骨髓基质干细胞培养 ,传代后改用含地塞米松、β -甘油磷酸纳和维生素C的条件培养基进行培养 ,用倒置显微镜观察、碱性磷酸酶染色、四环素荧光标记等方法对所培养的细胞进行生物学特性研究。　结果　接种细胞 4d即可传代 ,在条件培养液中 2周形成多层结构 ,并出现黑色结节 ,ALP染色阳性 ,结节四环素荧光标记、钙染色阳性 ,Ι型胶原免疫组化染色呈黄褐色。　结论　采用本法在体外培养的人成骨细胞生长良好 ,并具有成骨细胞形态特征和生物学特性 ,可做为临床所需的成骨种子细胞来源 ,为骨组织工程学研究奠定了实验基础。Objective To study the culture of human bone marrow stromal cells and its osteogenic characteristics in vitro, and provide osteoblast seed cells for bone tissue engineering. Methods Human bone marrow stromal cell obtained from human iliac crest was cultured in medium. The cells were passaged and were subcultured in media containing dexamethaseone, β-glycerphosphoric sodium and ascorbic aicd. During culture, the cells were examined by phase-contrast microscope , ALP strain, typeⅠcollagen strain, tetracycline fluorescence label. Results The stromal cells could be passaged in 4 days, and they transformed into multilayer structure in 2 weeks,and gradually congregated into diffused opaque clusters later in condition medium.ALP strain and tetracycline fluorescence label showed postive reaction,procollager typeⅠpropeptides were detectable. Conclusions The human bone marrow stromal cells cultured in this experiment survived well, the morphological and biological characteristics of the culture cells were similar to those of osteoblasts,the human bone marrow stroma cells cultured with conditions may offer a source of the osteogenous seed cells for clinical requirment.

关 键 词：骨髓基质干细胞 成骨细胞 细胞培养 人 

分 类 号：R-33[医药卫生]