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

• Technique and method • Previous Articles     Next Articles

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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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