成人骨髓成骨细胞体外培养

In Vitro Culture of Adult Human Osteoblasts from Bone Marrow

目的 研究成人骨髓基质干细胞在体外培养条件下的成骨能力及生物学特性 ,探讨简便易行的成人成骨细胞体外培养方法 ,为骨组织工程选择理想的种子细胞来源。方法 因诊断需要抽取健康成人骨髓组织 ,在含体积分数为 10 %胎牛血清的DMEM培养基中 ,置于 37℃、含体积分数为 5 %的CO2 湿化空气孵箱中培养 ,传代后改用含地塞米松、β-甘油磷酸钠和维生素C的条件培养基培养 ,用倒置相差显微镜观察、HE染色、扫描与透射电镜观察增殖和分化情况 ,并测定培养细胞的生长曲线 (MTT法 )、碱性磷酸酶活性和钙结节形成能力。结果 采用本法在体外培养的成人成骨细胞生长良好 ,生化指标稳定 ,表现出与典型的成骨细胞相似的形态特征和生物学特性。结论 本实验方法取材方便 ,所培养的细胞具有成骨细胞的特性 。

Objective To study osteogenic capability of adult human bone marrow stromal cells and biological characteristical identification in vitro in order to explore the appropriate methods of adult human osteoblast culture in vitro and to select an idea source of seed cells for bone tissue engineering.Methods Bone marrow was aspirated from the healthy adult human and cultured in DMEM containing volumes of 10% fetal bovine serum. The cells were incubated at 37℃ and relative humidity and in 5% CO 2 environment. Subsequently, dexamethasone, beta sodium glycerophosphate and ascorbic acid were added into culture medium to feed their passages. Proliferations and differentiations of all the cultured cells were observed continually under an inverted phase contrast microscope, a light microscope with HE stained,and a scanning and transmission electron microscope. Growth curve of the cultured cells was evaluated by MTT, activities of alkaline phosphatase and abilities of formation of calcifying nodules were detected.Results The adult bone marrow stromal cells cultured in this experiment survived well, the biochemical indexes were stable, and the cultured cells demonstrated a biological and morphologic characteristics similar to those of osteoblasts.Conclusion To draw the materials from bone is convenient in this experiment, and the cultured osteoblasts shows osteoblastic property, and can be used as a routine method of culturing seed cells of bone tissue engineering.