小鼠破骨细胞骨髓诱导培养体系的建立

Culture of osteoclasts induced from bone marrow mesenchymal stem cells in mice

目的:建立一个有效的破骨细胞培养体系,为研究和防治骨溶解和骨质疏松症提供实验基础。方法:在无菌条件下取小鼠股骨、肱骨,收集骨髓细胞,分为对照组和实验组。对照组不加入细胞因子;实验组加入巨噬细胞集落刺激因子(M-CSF)25 ng/m l和骨保护素配体(RANKL)25 ng/m l,进行破骨细胞骨髓诱导培养。观察比较两组抗酒石酸酸性磷酸酶(TRAP)染色阳性细胞和骨吸收陷窝,并用扫描电镜观察骨吸收陷窝形态。结果:实验组破骨细胞数量多,功能活跃,与对照组比较,差异有非常显著性意义(P<0.01)。结论:本实验成功建立了小鼠破骨细胞骨髓诱导培养体系。

Objective：To investigate a effective culture method of osteoclasts induced by bone marrow mesenchymal stem cells. Methods：Femurs and humerus were dissected and bone marrow cells were collected and cultured from 4-week-mice. Two culture methods according to the medium with macrophage colony-stimulating factor（M-CSF） at 25 ng/ml and receptor activator of nuclear factor-kB ligand （RANKL） at 25 ng/ml or not, were compared. The tartrate-resistant acid phosphatase（TRAP） positive multineucleated and bone resorption pits were counted, the pits of bone resorption was observed by scaning electronmicroscopy. Results：TRAP positive multinuclear cells were induced from mice bone marrow mesenchymal stem cells with both M-CSF and RANKL , and these cells formed resorption pits on bone slices. The M-CSF and RANKL had positive effect on growth of osteoclasts and bone resorption pits. Conclusion：The culture system of osteoclasts induced from bone marrow mesenchymal stem cells in the presence of M-CSF and RANKL was availabe established.