两种方法培养大鼠破骨细胞的噬骨能力比较  

Comparison of bone absorption capacity of rat osteoclasts cultured by two methods

作　　者：王正东[1] 颜南[1] 潘峰[2] 杨芳莉[1] 刘自力[1] 姜海波[1] 臧晋[1] 牟军[1] 柏树令[2] 

机构地区：[1]沈阳医学院,辽宁省沈阳市110034 [2]中国医科大学,辽宁省沈阳市110001

出　　处：《中国组织工程研究》2013年第20期3611-3617,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:原代培养的破骨细胞数量少,而诱导培养产生的破骨样细胞数量多,能够满足一些研究骨代谢实验的要求。但是两种细胞在特异酶及噬骨能力方面是否具有相同的效果,到目前为止,还没有详尽的数据来支持。目的:比较大鼠原代分离的破骨细胞及诱导形成的破骨样细胞噬骨能力上的差异。方法:取24h内新生Wistar大鼠四肢长骨,酶消化法分离培养破骨细胞,将破骨细胞培养过程中消化下来的骨髓单核细胞加入1,25(OH)2D3诱导生成破骨样细胞。苏木精-伊红染色、抗酒石酸酸性磷酸酶(TRAP)染色鉴定。用甲苯胺蓝染色共培养骨片比较两组破骨细胞噬骨能力。结果与结论:诱导第9天,破骨样细胞数目为破骨细胞数目的11倍,其形态与原代消化获得的细胞形态相同。抗酒石酸酸性磷酸酶染色呈阳性。甲苯胺蓝染色显示两组破骨细胞在骨片上培养时产生骨陷凹面积及深度差异无显著性意义。结果显示破骨样细胞的形态、特异酶及噬骨能力与破骨细胞无差异。BACKGROUND： There are less primary cultured osteoclasts, but a large number of osteoclast-like cells produced by induced culture, which is able to meet the requirements of some experimental studies of bone metabolism. But there are no detailed data to identify whether the osteoclasts and osteoclast-like cells have the similar effect on specific enzymes and bone absorption capacity. OBJECTIVE： To compare the difference of bone absorption ability between the primary cultured osteoclasts and induce cultured osteoclast-like cells from the rats. METHODS： The long bones were obtained from the limbs of 24 hours newborn Wistar rates, and the enzyme digestion was used to isolate and culture osteoclasts; then bone marrow mononuclear cell s separated from the osteoclasts during culture were added with 1,25（OH） 2 D 3 RESULTS AND CONCLUSION： The number of the osteoclast-like cell s was 11 times that of osteoclasts at 9 days after induction. The morphology of osteoclast-like cell s was similar with that of the primary cultured osteoclasts. The osteoclast-like cell s were positive for tartrate-resistant acid phosphatase. Toluidine blue staining results showed there was no significant difference in bone lacuna area and depth produced by the osteoclasts cultured on the bone slices between two groups. These findings show that there are no significant differences in morphology, specific enzymes and bone absorption capacity between osteoclast-like cell s and osteoclasts. to induce to generate the osteoclast-like cell s. The cell s were identified with heamtoxylin-eosin staining and tartrate-resistant acid phosphatase staining. The bone absorption ability of the osteoclasts in two groups was compared with toluidine blue staining.

关 键 词：组织构建 骨组织构建 破骨细胞 培养 诱导 鉴定 噬骨 1 25(OH)2D3 破骨样细胞 单核细胞 WISTAR大鼠 

分 类 号：R318[医药卫生—生物医学工程]