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目的 :探讨简易的成人成骨细胞体外培养方法 ,为进一步研究成骨细胞及骨组织工程提供技术平台。方法:手术中抽取健康成人骨髓组织 ,采用全骨髓培养法在体外进行培养 ,细胞汇合后改用含地塞米松、β-甘油磷酸钠和维生素 C的条件培养基进行传代培养 ,观察细胞增殖和分化情况 ,并对细胞进行鉴定。结果:培养的成人成骨细胞表现出成骨细胞的形态特征 ,细胞碱性磷酸酶 (AL P)阳性率为 6 8% ,经 Von Kossa’s染色证实其能最终形成钙结节。结论:全骨髓培养法所培养的细胞具有成骨细胞的特性 ,可作为骨组织工程种子细胞培养的常规方法。
OBJECTIVE: To explore a simple method for culturing adult osteoblasts in vitro and to provide a technological platform for further research on osteoblasts and bone tissue engineering. Methods: Bone marrow tissues from healthy adults were collected and cultured in vitro by whole bone marrow culture. After the cells were confluent, the conditioned medium containing dexamethasone, β-glycerophosphate and vitamin C was used for subculture, and the cell proliferation and Differentiation, and identification of cells. RESULTS: The cultured adult osteoblasts showed morphological characteristics of osteoblasts. The positive rate of alkaline phosphatase (ALP) was 68%. It was confirmed by Von Kossa’s staining that it eventually formed calcium nodules. CONCLUSION: The cells cultured by whole bone marrow culture have the characteristics of osteoblasts and can be used as routine methods for culturing bone tissue engineering seed cells.