[Purification and study of the characteristics of the multinucleated giant cells of the giant cell tumor].
OBJECTIVE
To harvest high purified osteoclast-like giant cells from the giant cell tumor by using trypsin and EDTA(ethylenediamineteraacetic acid) digestion. And explore the role and the origin of the multinucleated giant cells of the giant cell tumor.


METHODS
Multinucleated giant cells were isolated and cultured from 8 cases of giant cell tumor of the long bone. After 20 hours of culture, 0.5 g.L-1 trypsin/0.2 g.L-1 EDTA was applied to detach the mononuclear stromal cells. The isolated multinucleated giant cells were inoculated on dentine slice to observe bone resorption ability in vitro. Immunohistochemistry study with V-ATPase, CA II (carbonic anhydrate II), cathepsin K, MMP-9 (matrix metalloproteinase-9), CD68 and TRAP(tartrate-resistant acid phosphatase) staining were carried out in the purified multinucleated giant cells.


RESULTS
We obtained 85% purified multinucleated giant cells by trypsin/EDTA digestion. Multinucleated giant cells formed resorption lacunae on dentine slice in vitro and positively stained for V-ATPase, CA II, cathepsin K and MMP-9. The TRAP staining was also positive.


CONCLUSIONS
Using 0.5 g.L-1 trypsin/0.2 g.L-1 EDTA digestion can get high purified multinucleated giant cells from the giant cell tumor. The multinucleated giant cells match all the character of the osteoclast and may originate from the fusion of the mononuclear stromal cells of the lesion.
Liu
Bo
B
Department of Oral Pathology, Peking University School of Stomatology, Beijing 100081, China. Liubodentist@sina.com
Yu
Shifeng
S
Pang
Shuzhen
S
D001859:Bone Neoplasms / Q000473:pathology*; D024402:Carbonic Anhydrase II / Q000032:analysis; D002469:Cell Separation / Q000379:methods*; D004492:Edetic Acid / Q000494:pharmacology; D018212:Giant Cell Tumor of Bone / Q000473:pathology*; D006801:Humans; D007150:Immunohistochemistry; D020780:Matrix Metalloproteinase 9 / Q000032:analysis; D010010:Osteoclasts / Q000737:chemistry / Q000166:cytology*
D004492:Edetic Acid; D020780:Matrix Metalloproteinase 9; D024402:Carbonic Anhydrase II