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A Simplified Method for Purifiing Osteoclasts from Human Giant Cell Tumor of Bone
Yun-Lin Wang*, Xiao-Qin Liu, Qin Xia, Yu-Hong Guan
Department of Gerontism, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
Abstract: 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 0.2% type I collagenase. We characterized OCs in terms of the expression of different phenotypic markers of OCs. the phenotypic markers of OC include Tartrate-resistant acid phosphatase staining (TRAP)， the expression of calcitonin receptor (CTR)， cathepsin K and receptor activator of necrosis factor kB (RANK) mRNA were examined by RT-PCR. The OCs cell purified by above method， the purity is about 79.7%. They showed the normal osteoclast phenotypes markers of OC. The method provided a system for performing biochemical and molecular studies of OCs. The study indicated that the method of purifying the osteoclasts from human giant cell tumor of bone cell could be used for research of bone metabolism.
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