ABSTRACT
Objective To establish a stable and useful method for culturing human osteoclast-like cells in vitro,and investigate the effect of 1?,25-(OH)2D3,M-CSF and PGE2 on osteoclasts differentiation,proliferation and activation so as to lay the foundation for further study of the biological mechanism for tooth movement.Methods The HCMNC were isolated and cultured in 24-well plate with coverslips and human dentine slices.The experiment group was cultured with 1?,25-(OH)2D3,M-CSF and PGE2,respectively,while the control group was not.The liquid was changed every 3 days and the whole culture process lasted for 7 days.The phase contrast microscopy and TRAP staining were adopted to identify osteoclast-like cells.Results On the 3rd day the monocytes began to fuse and on the 7th day positive multinucleated cells could be seen with TRAP staining,but absorption pit was not formed on the dentin slices.The group with 1?,25-(OH)2D3 had the largest number of osteoclast-like cells.Conclusion After the monocytes in UCB are cultured by 1?,25-(OH)2D3,M-CSF,PGE2 induction,they can turn into TRAP(+) multinucleate osteoclast-like cells,the 1?,25-(OH)2D3 10-8mol/L being the most effective.