人胚成骨细胞的分离培养及生物特性的研究

目的 观察体外条件下成骨细胞生物学特性，为研究骨代谢提供一种体外模型。方法取人胚胎颅骨骨膜，采用酶消化法获取成骨细胞体外培养并传代。观察细胞形态，生物特性，绘制生长曲线，并经碱性磷酸酶染色鉴定成骨细胞以及比较冻存前3～5代与冻存后成骨细胞的特点。结果体外培养的骨膜成骨细胞形态多为梭形和多角形与成纤维细胞相似，3～5代成骨细胞生长特点稳定，具有成骨能力，冻存后成骨细胞生存率、增殖能力明显受到影响。结论自骨膜获取人成骨细胞培养方法易形，可大量培养高纯度的人成骨细胞供研究使用，冻存细胞不易作为研究对象。

Separating Cultivating and Growth Feature Study of Human Embryo Osteoblast

Objective To observe osteoblasts creature feature in vitro and provide a kind of method for researching bone metabolism. Methods The embryo calvarial periosteum tissue was taken and, human osteoblasts were obtained by enzyme-assimilating methods. The morphological change, growth feature and osteogentic capability of osteoblasts were observel during culture in vitro, drew the growth curve was graphed and the cells was identified by alkaline phosphatase dye. At the same time, the morphology and bioactivity of 3to 5 th-generation osbeoblasts and anabiotic cells were studied comparatively. Results Most of the cultivated periosteal-derived osteoblasts were polygon-like or shullfle-like similar to fibroblasts and the 3 to 5 th generation osteoblasts had the stable growth feature and osteogenetic capability. Low temperature affected the surrival rate and proliferation of osteoblasts after cryopreservation. Conclusion The cultivated periosteal-derived osteoblasts were matured ones which growed and breed rapidly and could become the more ideal seeded cells for study. The osteoblasts after cryperservation were not fit for the research.