上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (10): 1303-.doi: 10.3969/j.issn.1674-8115.2010.10.029

• 技术与方法 • 上一篇    下一篇

钛合金颗粒存在下破骨细胞的体外诱导培养

毛远青, 朱振安, 汤亭亭, 袁 振, 王晓庆, 刘 铭   

  1. 上海市骨科内植物重点实验室 上海交通大学 医学院附属第九人民医院骨科, 上海 200011
  • 出版日期:2010-10-25 发布日期:2010-10-27
  • 通讯作者: 朱振安, 电子信箱: zhuzhenan2006@126.com。
  • 作者简介:毛远青（1974—）, 男, 主治医师, 博士；电子信箱: maoyuanqing@yahoo.cn。
  • 基金资助:

    上海市自然科学基金（08ZR1413000）；上海市骨科内植物重点实验室建设专项经费（08Dz2230300）；上海教委重点学科建设基金（J50206）

In vitro induction and culture of osteoclasts with Ti-alloy particles

MAO Yuan-qing, ZHU Zhen-an, TANG Ting-ting, YUAN Zhen, WANG Xiao-qing, LIU Ming   

  1. Shanghai Key Laboratory of Orthopaedic Implant, Department of Orthopaedics, The Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
  • Online:2010-10-25 Published:2010-10-27
  • Supported by:

    Shanghai Natural Science Foundation, 08ZR1413000；Shanghai Key Laboratory of Orthopaedic Implant Foundation, 08Dz2230300；Shanghai Education Committee Foundation, J50206

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摘要：

目的 探讨钛合金颗粒存在下小鼠巨噬细胞体外诱导培养为成熟破骨细胞的方法。方法 获取小鼠骨髓巨噬细胞，采用巨噬细胞集落刺激因子依赖性前体细胞诱导法，于钛合金Ti-6Al-4V颗粒存在条件下进行诱导培养，培养后细胞行抗酒石酸酸性磷酸酶（TRAP）染色，RT-PCR法检测破骨细胞特异性TRAP、CK、CR mRNA表达；采用相同方法于骨片上诱导培养细胞并观察骨吸收陷窝。结果 Ti-6Al-4V颗粒存在下培养第6天细胞开始出现融合，第9天出现多核巨细胞，TRAP染色阳性，细胞内有破骨细胞特异性TRAP、CK、CR mRNA表达；于骨片上诱导培养的细胞可形成骨吸收陷窝。结论 钛合金颗粒存在下，成功将小鼠骨髓巨噬细胞体外诱导培养为成熟破骨细胞。

关键词: 巨噬细胞, 破骨细胞, 诱导培养, 颗粒

Abstract:

Objective To explore the method of in vitro induction and culture of mature mouse osteoclasts from macrophages with Ti-alloy particles. Methods Mouse macrophages were obtained from bone marrow, and macrophage colonystimulating factor (M-CSF)-dependent precursor method was employed for cell induction and culture with Ti-6Al-4V (Ti-alloy) particles. The cultured cells subjected to tartrate-resistant acid phosphatase (TRAP) staining, and expressions of osteoclast specific TRAP mRNA, CK mRNA, and CR mRNA were detected by RT-PCR. Cells were induced and cultured on bone flaps with the same method, and absorption lacuna was observed after culture. Results Cells began to merge on the sixth day after culture with Ti-6Al-4V particles, and multinucleated giant cells emerged on the ninth day, with positive TRAP staining and expressions of osteoclast specific TRAP mRNA, CK mRNA, and CR mRNA. Absorption lacuna formed on bone flaps with cultured cells. Conclusion Mature mouse osteoclasts can be successfully induced and cultured in vitro from macrophages with Ti-alloy particles.

Key words: macrophage, osteoclast, induction and culture, particle