摘要
研究了鸡破骨细胞分化因子(Chicken osteoclast differentiation factor,chODF)体外诱导鸡骨髓细胞形成破骨样细胞的能力。在无菌条件下取鸡股骨和胫骨,收集骨髓细胞。试验分设A、B、C、D、E、F、G 7组。A组为对照组,不加细胞因子,其他各组为试验组,其中B组仅加chODF,C组只加巨噬细胞集落刺激因子(Macrophage-colony-stimulating factor,M-CSF),D、E和F组同时加不同浓度的chODF和M-CSF,G组同时加chODF、M-CSF和鸡护骨素(Chicken osteoprotegerin,chOPG)。通过抗酒石酸盐酸性磷酸酶(Tratrate-resistant acid phosphatase,TRAP)、HE、骨吸收陷窝甲苯胺蓝染色和显微、超微检查,观察和鉴定破骨样细胞(Osteoclast like cell,OLC)的形成。结果表明,chODF可诱导形成典型的OLC,骨吸收陷窝清晰可见,其陷窝面积大,数量多,且TRAP阳性多核细胞的数目与ODF的浓度呈正相关,多组比较其差异显著(P<0.05),但若加入chOPG,则阻断了OLC的形成和骨吸收。本试验建立了一种鸡骨髓细胞诱导破骨样细胞方法,既为体外研究鸡破骨细胞的分化发育和功能调节创造了条件,也为进一步研究OPG/ODF/RANK(NF-κB受体,Receptor activator of NF-κB)在蛋鸡骨质疏松症等骨骼疾病的作用机制提供理论基础。
The purpose of this study was to investigate an effective culture method of osteoclasts （OC） formation and to test the possibility that chODF （chicken osteoclast differentiation factor） was sufficient for OC development in primary chicken embryo marrow cell culture. Femurs and humerus were dissected and bone marrow cells were collected and cultured in seven culture methods according to the medium with different concentration of chODF, M-CSF and chOPG. OC formation was observed and indentified by TRAP （tartrate-resistant acid phosphatase）, HE, toluid- ine blue staining, phase-contrast microscope and scanning electron microscopy. Results showed that the combination of chODF and M-CSF could induce OLCs formation and bone resorption in chicken embryo marrow cell culture. The number of TRAP-positive multi-nucleated cells in the culture medium increased dose-dependently with ODF concentration （P〈0.05）, at meanwhile, OPG can interrupt the formation of OLC. In conclusion, we established an effective way to form OLC, and it could be employed to investigate the differentiation and activation of OC. It also provide theory of mechanism of OPG/ODF/RANK in the process of osteoporosis in layer hens.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2008年第8期1127-1131,共5页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(30671546)
教育部重点项目(107059)
江苏省自然科学基金(BK2004099)
高校博士点基金项目(2004037036)
