Osteoclasts, which are responsible for bone resorption, are rare cells with only 2-3 cells seen per 1 mm3 of bone. However, the loss of function in osteoclasts, problems with their differentiation and decrease in their number lead to bone osteosclerosis/osteopetrosis. On the other hand, an increase in their number or function induces bone osteoporosis, indicating that osteoclasts play a pivotal role in bone homeostasis. It has been demonstrated that bone destruction and hypercalcemia induced by metastatic tumors are carried out by osteoclasts activated by the tumor cells, and the inhibition of osteoclast formation prevents the bone destruction and even bone metastasis. Abnormal osteoclast function is closely related to various diseases. Furthermore, osteoclasts are indispensable in forming bone marrow to produce blood cells, and the absence of osteoclasts causes osteopetrosis, resulting in extramedullary hematopoiesis. Although the physiological roles of osteoclasts are well described, the mechanisms of their differentiation remain to be elucidated. Recently, RANK (receptor activator of nuclear factor kappaB) and its ligand (RANKL) have been identified and their essential roles in osteoclastogenesis have been demonstrated, which has provided new insights into the osteoclast differentiation pathway. We have established an in vitro osteoclast culture system by isolating osteoclast precursor cells and culturing them in the presence of macrophage colony stimulating factor (M-CSF) and soluble RANKL. This system has enabled us to analyze the regulation mechanisms in osteoclast formation.