番鸭破骨细胞的培养及鉴定

Culture and identification of osteoclasts from the duck

分离7日龄内番鸭长骨破骨细胞(osteoclast,OC),倒置显微镜观察其活体形态。培养1、3、5、7d进行抗酒石酸酸性磷酸酶染色(TRAP),培养7d后进行骨吸收陷窝定量分析,观察番鸭OC数量、体外生存时间以及骨吸收功能。结果显示,倒置显微镜下观察,OC为多核巨细胞,有伪足并借助伪足的凹凸伸缩而改变形态、运动行走。玻片上OCTRAP染色阳性,培养7d的象牙片上产生明显吸收陷窝。直接分离OC的数量随培养时间延长而减少,单核细胞融合而成的OC数量随培养时间延长而增加;分离培养的鸭OC具有骨吸收活性,TRAP染色阳性。

Establishing the method of culture and identification of osteoclastic cells from long hones of duck,to provide the research foundation of various factors contributing to hone malnutrition. Osteoclasts were isolated from duck （ 1-7 days） long hones. The shape and function of these osteoclasts were observed and identified by phase-contrast microscope, TRAP staining, Toluidine blue staining. Both morphological and functional studies showed that the isolated cells share some of the typical characteristics of osteoclasts： multinuclearity; developmental spreading and pseudopodial activity when cultured on glass;high tartrate-resistant acid phosphatase （TRAP） ;resorption lacunae could he found when the cells were co-cultured with devitalized bone slices and the number increased as the time followed. The number of degraded osteoclasts decreased along with the time of culture, while the number of forming osteoclasts of monocytes increased. The osteoclasts could make resorption lacunae on the dentine discs.