何涛 杨庆铭 邓廉夫 200025 上海瑞金医院上海市伤骨科研究所 中华骨科杂志 1999 0 19 1


    关键词：骨髓；破骨细胞；细胞分离；细胞；培养的 期刊 zhgkzz 0 实验研究 fur -->


    

【摘要】目的 在国内前人建立动物破骨细胞骨髓培养方法的基础上，首次建立人破骨细胞骨髓培养法。方法 采取13例25～75岁行全髋置换术的成人股骨上端红骨髓，经密度梯度离心得骨髓单个核细胞(marrowmononuclear cells,MMNC)，在浓度为1×10^-8 mol/L的1,25-(OH)₂ D₃ 的促分化作用下，于预置盖玻片或人胫骨皮质骨片的24孔培养板内，每孔接种1×10⁶ /ml培养1～3周。结果 光镜下观察有单个核细胞融合成多核细胞和单个核细胞团形成，经TRAP染色呈阳性反应，电镜下观察有典型的骨吸收陷窝形成。结论 这些均表明由骨髓培养破骨细胞样细胞的方法是成功的。而且利用此方法较之新生动物的直接分离法，所得破骨细胞量多，特征明显，并且方法简单易行。更重要的是为人的破骨细胞样细胞，因此培养方法更有价值。

    The Establishment andIdentification of Human Adult Osteoclast Marrow Culture Method

HE Tao, YANG Qingmimg, DENGLianfu.

    Ruijin Hospital and Shanghai Institute of Traumatology and Orthopaedics, Shanghai 200025

【 Abstract 】 Objective Toestablish a method for human osteoclast marrow culture on the base of previously publishedmethod. Mothods Red marrow in the upper femur was procured from 13 patients (aged 25～ 75 years) during the total hipjoint replacement procedures. The specimen was treated with Ficoll- Hapaque centrifuge to get MMNC which was then, culturedunder the 1×10^-8 mol/L 1,25- (OH)₂ D₃ differentiating effect in 1×10⁶ cells/ml per well on the 24 wellculture plates with bone slices or cover- slips for 1～ 3 weeks. Results Mononuclear cells fusedto form multinuclear cell clusters in micrograph. They were postive in TRAP staining andcould form typical resorption pits in SEM. Conclusions The result provedthat the marrow culture method was successful ......