天津医药

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RAW264.7细胞在体外分化成破骨细胞的研究

程明1,戴闽2,张斌3,程细高3   

  1. 1. 南昌大学第一附属医院骨二科
    2. 南昌大学第一附属医院
    3.
  • 收稿日期:2010-01-15 修回日期:2010-03-15 出版日期:2010-07-15 发布日期:2010-07-15
  • 通讯作者: 程细高

The Study of RAW264.7 Cells Differentiate into Osteoclasts in Vitro

  • Received:2010-01-15 Revised:2010-03-15 Published:2010-07-15 Online:2010-07-15

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摘要： 摘要：目的：观察小鼠的单核巨噬细胞RAW264.7的一般生物学特征及验证在RANKL（重组细胞核因子κB受体活化因子配基，Ligand of receptor activator of NFκB)诱导下形成成熟的有骨吸收能力的破骨细胞的可行性。方法：RANKL诱导细胞后，用TRAP (抗酒石酸酸性磷酸酶)染色法观察阳性多核细胞，扫描检测电镜骨吸收陷窝。结果：RANKL诱导RAW264.7第3天开始出现少数多核巨细胞，随时间的延长细胞数目逐渐增多，7天多核巨细胞数达峰值，多核巨细胞TRAP染色阳性，电镜下可见骨片上的吸收陷窝呈圆形或椭圆形，边界轮廓清晰，陷窝底部纤维腐蚀吸收的纹路清晰可见，提示多核巨细胞具有骨吸收功能。结论：RAW264.7是一种较好的破骨前体细胞模型。RAW264.7诱导形成的多核巨细胞具有破骨细胞特征，是可用于实验研究的成熟破骨细胞。RANKL诱导RAW264.7形成破骨细胞方法简便易行、可重复性好。

关键词: 核因子κB受体活化因子配基, RAW264.7细胞, 破骨细胞, 抗酒石酸酸性磷酸酶染色

Abstract: Abstract Objective:To observe the biologic characteristics of RAW264.7 cells,and to validate the feasibility of RANKL-mediated differentiation of RAW264.7 cells into bone-resorptive Osteoclasts (OCs).Methods：RAW264.7 cells were treated with RANKL. RANKL-induced formation into OCs were observed after tartrate resistant acid phosphatase(TRAP) staining. Bone resorption pits were detected by scanning electron microscope. Results：RANKL-differentiated RAW264.7 cell cultures develop both TRAP+ mononuclear and multinucleated cells.It has the ability of generating bone resorptive pits. Conclusions：RANKL can induce RAW264.7 cells to mature OCs. RAW264.7 cells are easier to obtain and culture.So RAW264.7 cells can be used as a preferable preosteoclast model.This methods has good repeatability and feasibility.It represent a relatively homogeneous population of pre-OCs.

Key words: ligand of receptor activator of NF-κB（RANKL）, RAW264.7 cell, osteoclasts, tartrate-resistant acid phosphatase（TRAP）stain