【作者】 白鹤；

【导师】 赵劲民； 杨志； 苏伟；

【作者基本信息】 广西医科大学 ， 骨外科， 2003， 硕士

【摘要】 目的：建立一种骨髓基质细胞(BMSCs)体外培养，定向诱导分化为成骨细胞的方法，并探讨其体外成骨能力。 方法：冲洗法获取SD大鼠骨髓，使用percoll分离液密度梯度离心分离提纯BMSCs，进行体外培养，保留贴壁细胞传代，在培养液中添加地塞米松(Dex)、维生素C(VitC)、β-甘油磷酸钠(β-GP)，观察细胞生长状态，通过碱性磷酸酶(ALP)染色和Von Kossa染色观察其向成骨细胞分化及体外成骨情况。 结果：BMSCs呈成纤维细胞样生长，增殖能力强，生长迅速。诱导条件下细胞的ALP活性明显增高，并且出现钙化结节。 结论：使用本实验方法，获得的BMSCs增殖能力强，在地塞米松、维生素C和β-甘油磷酸钠联合诱导下可定向分化为类成骨细胞，成骨能力肯定，适合作为骨组织工程的种子细胞。更多还原

【Abstract】 Objective To establish a method to culture the bone marrow stromal cells (BMSCs) from rats and induce them to osteoblasts in votro, and to explore its osteogenetic ability.Methods The bone marrow was aspirated from the bone of limbs of rats and was isolated by gradient centrifugation in percoll. The mononuclear cells were collected and cultured in vitro, and the adherent cells were kept. The higher purity of BMSCs were obtained by repeatedly removal of nonadherent cells. During the passage, BMSCs were cultured in the defined medium containing the dexamethasone, VitC and Natrium- -glycerophosphat and the growth and proliferation were observed . The differentiation to osoblasts andosteogenetic conditions in vitro were observed by the staning of ALP and Von Kossa.Results BMSCs grew rapidly and proliferated in visible symmetric colonies with fibroblast appearance. The ALP activity of passage cells increased dramatically and the calcification nodules occured under the indecement.Conclusion BMSCs were obtained and showed the strong ability of proliferation by using this method, and in the define medium were induced and differentiated to fibroblasts with Dex, VitC and -PG. So BMSCs can be served as the seeding cells in bone tissue engineering.更多还原