Comparison of the Culturing Methods of Osteoclasts between Induced and Mechanical Procedure

Objective To investigate the differences of morphology and bone absorption function of osteoclast(OC)between different cultural methods,and to provide the basis for further investigation on the OC in vitro.Methods Mechanical separation method was used to isolate mature osteoclasts from long bones of SD rats aged one-day,and induced culture method was applied to induce osteoclast-like cell(OLC) by using RANKL(100 ng/mL)and M-CSF(100 ng/mL).The morphological and functional changes of OC as well as OLC were observed.Results OC and OLC were multinucleated giant cells,which could be stained positively by TRAP,and could form bone absorptive lacuna.The number of OLC in inducement method was more than in mechanical separation method,but bone absorption lacuna in mechanical separation method was smaller and shallower as compared with inducement.Conclusions OC derived directly from long bones is less in number,but is more active in bone absorption,thus can be appropriate for the study of bone absorption,migration and adhesion,apoptosis and monocellular molecular biology.OLC obtained by inducement method is more in number,but less active in bone resorption and is better for the study of OC differentiation and development.