人骨膜源性成骨细胞的体外培养

目的：探索成骨细胞的体外培养方法，建立人骨膜源性成骨细胞体外实验模型。方法；以人骼骨骨膜为材料，通过植块培养法进行细胞培养，通过形态学观察；细胞的碱性磷酸酶（ＡＬＰ）染色；培养液中骨钙放免分析等进行细胞学鉴定。结果：成骨细胞为成纤维细胞型，，贴壁生长，形态多样化，ＨＥ染色细胞轮廓清晰，核内可见２～３个核仁；细胞的ＡＬＰ染色淡了性；培养液中检测出较高浓度的ＢＧＰ。结论：可以利用人骨膜灯民骨细胞的体外

THE CULTURE OF PERIOSTEAL-DERIVED OSTEOBLAST (POB) IN VITRO

Objective: To explore the culture of periosteal-derived osteoblast (POB) in vitro to establish an experimental model for the study of human bone metabolism, growth control and environmental effects. Methods: Human osteoblasts were obtained by explanting the iliac periostenum culture in DMEM media. The cytomorphological observation, the cytochemical staining of AKP and the detection of BGP (Bone Gla Protein or osteocalcin) in supernate were carried out to identify the osteoblast. Results: The results of the cell culture in vitro and the cytologic identification show: (1) The periosteal-derived osteoblast (POB), a fibroblast typed cell which grows by adhering to the culture flask, is spindle-shaped with 2-3 nucleoli in an oval nucleus. It has a few particulate matters in its transparent cytoplasm; (2) The positive rate of osteoblast stained by AKP is over 80%. (3) The higher concentration of BGP can be detected in its supernate, while there is little BGP in DMEM medium without osteoblast culture. Conclusions: Human osteoblast can simply reliably be isolated and culture from periostenum in vitro.