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利用破骨细胞贴壁快以及耐胰蛋白酶地特性,采用0.25%胰蛋白酶和0.2%I型胶原酶来分离纯化骨巨细胞中的破骨细胞,即得到破骨细胞,并进行细胞学和分子生物学鉴定,包括抗酒石酸酸性磷酸酶染色和HE染色,并采用RT-PCR反应方法检测降钙素受体、组织蛋白酶K和破骨细胞分化因子受体的表达。该方法所得细胞纯度可达79.7%,具有破骨细胞表型特征。可用于生化和分子生物学研究,是进行骨细胞学研究的破骨细胞来源。
Using osteoclast adherent fast and trypsin-resistant properties, osteoclasts were isolated and purified from bone giant cells using 0.25% trypsin and 0.2% type I collagenase to obtain osteoclasts and cytology and molecules Bioassay, including tartrate-resistant acid phosphatase staining and HE staining, and the expression of calcitonin receptor, cathepsin K and osteoclast differentiation factor receptor were detected by RT-PCR. The purity of the obtained cells is up to 79.7%, which has the characteristic of osteoclast phenotype. Can be used for biochemical and molecular biology research, osteoclast research osteoclast source.