A Simplified Method for Purifying Osteoclasts from Human Giant Cell Tumor of Bone

Abstract

Objective: To purify and identify the osteoclasts from the tissue of human giant cell tumor of bone. Methods: We have developed a new method that allows the purification of large numbers of authentic osteoclasts (OCs). The OCs were isolated from tissue of human giant cell tumor of bone by 0.25% trypsin and collagenase. We characterized OCs in terms of the expression of different phenotypic markers of OCs. The phenotypic markers of OC included Tartrate-resistant acid phosphatase staining (TRAP). The expression of calcitonin receptor (CTR), cathepsin K and receptor activator of necrosis factor κB (RANK) mRNA were examined by RT-PCR. Results: The OC cell purified by above method functioned normally in vitro. The purity was about 79.7%. They showed the normal osteoclast phenotypes markers of OC. Conclusion: The method provides a system for performing biochemical and molecular studies of OCs. The study indicates that the method of purifying the osteoclasts from human GCT cell can be used for research of bone metabolism.