Liu Bo, Yu Shifeng, Pang Shuzhen
Department of Oral Pathology, Peking University School of Stomatology, Beijing 100081, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2003 Feb 18;35(1):23-7.
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.
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.
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.
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.
采用胰蛋白酶和乙二胺四乙酸（EDTA）消化法从骨巨细胞瘤中获取高纯度破骨样巨细胞，探讨骨巨细胞瘤中多核巨细胞的作用及起源。
从8例长骨骨巨细胞瘤中分离培养多核巨细胞。培养20小时后，用0.5g.L-1胰蛋白酶/0.2g.L-1 EDTA分离单核基质细胞。将分离出的多核巨细胞接种于牙本质片上，观察其体外骨吸收能力。对纯化的多核巨细胞进行V-ATP酶、碳酸酐酶II（CA II）、组织蛋白酶K、基质金属蛋白酶-9（MMP-9）、CD68和抗酒石酸酸性磷酸酶（TRAP）染色的免疫组织化学研究。
通过胰蛋白酶/EDTA消化法获得了85%纯度的多核巨细胞。多核巨细胞在体外牙本质片上形成吸收陷窝，V-ATP酶、CA II、组织蛋白酶K和MMP-9染色呈阳性。TRAP染色也呈阳性。
使用0.5g.L-1胰蛋白酶/0.2g.L-1 EDTA消化法可从骨巨细胞瘤中获得高纯度的多核巨细胞。这些多核巨细胞符合破骨细胞的所有特征，可能起源于病变单核基质细胞的融合。
