鸡破骨细胞分离培养方法的建立

The establishment of a method of isolated and cultured chicken osteoclasts

为了寻找一种获得大量、纯化、有活力的破骨细胞的方法，我们对已经建立的兔破骨细胞体外分离培养方法的基础上加以改进。利用冲洗的方法，从２８天的鸡的长骨中得到细胞团１破骨细胞后，又对鸡的长骨相继进行胶原酶和胰蛋白酶的消化，得到细胞团２和细胞团３破骨细胞。将这些分离的细胞与盖玻片或骨磨片共同培养，通过相差显微镜观察到：这些分离的多核巨细胞能够运动，并能在骨磨片上形成吸收陷窝。另外，这些细胞对酸性磷酸酶染色呈阳性反应，而酸性磷酸酶是鉴别破骨细胞的标志。表明此分离培养鸡破骨细胞的实验技术是成功的。通过此方法获得的鸡破骨细胞比用以往方法获得的兔破骨细胞量多，且活力强。本方法的建立，为进一步研究骨吸收机理奠定了基础。

In this study, in order to obtain large numbers of pure and viable osteoclasts, we improved the method of isolated and cultured rabbit osteoclasts. The osteoclastic population 1 were obtained from 28-day old chicken long bones by flushing the marrow cavities repeatedly. The bones were then treated successively with collagenase and trypsin. The osteoclastic population 2 and population 3were obtained. These isolated cells were cultured on glass of bone slices. Under phase-contrast microscope, it has been found that these isolated multinucleated giant cells can migrate and generate resorption lacunae on bone slices. In addtion, these cells stained positively for acid phosphatase which aremarker for osteoclasts. These results suggested that the procedure of isolation of osteoclasts from thelong bones of chicken was successful. The numbers of chicken osteoclasts are larger obtained by this method than rabbit osteoclasts obtained by old method. The vitality of the chicken osteoclasts are stronger. This method lays a foundation for further study of pathogenesis of bone resorption.