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目的 :建立体外分离、培养兔和人成骨细胞的生物学模型 ,并对其生物学特性进行观察。方法 :抽取新西兰兔骨髓 ,采用全骨髓法培养 ;人髂骨松质骨经胰酶消化获得成骨细胞 ,以 1× 10 6 / ml的细胞浓度进行培养 ,约10 d左右得到贴壁生长的单层细胞。随后传代培养 ,对所获得的细胞进行生物学特性观察。 结果 :获得的细胞呈多种形态 ,有长短不一、粗细不均、互相连接的胞浆突起 ,细胞可相互重叠呈复层生长 ,并形成钙结节。经连续传代 ,细胞形态与功能不变 ,具有典型成骨细胞的生物学特性。结论:无论采用兔骨髓基质细胞全骨髓法 ,还是人松质骨经胰酶消化法均能在体外培养出大量高纯度成骨细胞 ,方法简便、易行 ,为成骨细胞复合生物降解材料移植修复骨缺损奠定了基础 ,而且培养的成骨细胞可作为生物学模型 ,供干预研究使用。
OBJECTIVE: To establish a biological model of rabbit osteoblasts and human osteoblasts isolated and cultured in vitro and to observe their biological characteristics. Methods: Bone marrow of New Zealand rabbits was harvested and cultured by whole bone marrow method. Osteoblasts were obtained from human iliac cancellous bone by trypsinization and cultured at a cell concentration of 1 × 10 6 / ml for about 10 days Monolayer cells. Subsequent subcultures were performed to observe the biological characteristics of the cells obtained. Results: The cells obtained in various forms, with varying lengths, uneven thickness, interconnected cytoplasm protrusions, cells can overlap each other was stratified growth, and the formation of calcium nodules. After continuous passage, cell morphology and function unchanged, with the typical biological characteristics of osteoblasts. CONCLUSION: A large number of high-purity osteoblasts can be cultured in vitro using whole-marrow bone marrow stromal cells of rabbit bone marrow stroma, or human cancellous bone by trypsin digestion. The method is simple and easy to perform, and it is a promising method for osteoblast composite biodegradable material transplantation Repair of bone defects laid the foundation, and cultured osteoblasts can be used as a biological model for intervention studies.