两种方法培养大鼠破骨细胞的比较研究

目的比较分析直接分离培养的Wistar大鼠破骨细胞（osteoclast，OC）和诱导培养的Wistar大鼠破骨样细胞（osteoclast-like cell，OLC）的形态和功能的差异，为体外药物干预试验奠定基础。方法采用两种培养法，即从新生（24h内）的Wistar大鼠的四肢长骨骨髓腔内壁机械分离成熟OC直接培养和10^-8mol／L的1，25（OH）2D3诱导4周龄Wistar大鼠骨髓单核细胞形成OLC的方法，对获得的OC／OLC进行形态和破骨功能观察。结果两种方法都培养出了抗酒石酸酸性磷酸酶（tartrate resistant acid phosphatase，TRAP）染色阳性的多核细胞，诱导法获得的破骨细胞数量较多（P〈0．05）。成熟OC与诱导获得的OLC形态特征相似，但后者在骨片上形成的陷窝较小而浅。结论直接分离培养法可获得骨吸收功能较活跃的OC，但数目较少，适合骨吸收功能分析、破骨迁移黏附、凋亡研究及单细胞分子生物学研究。1，25（OH）2D3诱导鼠骨髓单核细胞形成的OLC数量较多，但骨吸收功能较差，适合用于破骨细胞分化发育过程的研究。

Comparative study on two different culturing methods of osteoclasts of rats

Objective To study the morphological and functional difference between osteoclast(OC) and osteoclast-like cells(OLC) cultured by different means and to provide the basis for further investigation on the regulation of bone resorption by OC. Methods OCs were isolated from the long bones of neonatal Wistar rats, and OLC were induced from the marrow mononuclear cells of 4-week-old of Wistar rats by 10 -8 mol/L 1,25(OH)_2VitaminD_3 in vitro. The morphology and resorption function were observed for OC and OLC. Results The tartrate resistant acid phosphatase (TRAP) positive polykaryocytes were obtained by both means, and the amount of OLC was more than that of OC (P<0.05). The appearance of OLC was similar to that of OC, but the resorption function of OLC was not as stronger as OC. Conclusions OC derived directly from long bones is less in number, but is active in bone resorption, thus can be appropriate for the study of bone resorption, migration and adhesion, apoptosis and monocellular molecular biology. The bone resorption ability of OLC obtained by inducing means is more in number, but less active and is better for the study of OC differentiation and development.