骨髓间充质干细胞诱导为成骨细胞的初步研究

目的观察兔骨髓间充质干细胞的生长特点及诱导条件下的成骨能力。方法抽取兔骨髓组织,梯度离心后,保留贴壁细胞传代,稳定传代后改用含地塞米松、β-甘油磷酸钠和维生素C的诱导培养液,通过倒置显微镜观察,碱性磷酸酶染色,骨连接素、骨桥素免疫细胞化学染色等方法进行观测。结果原代兔骨髓间充质干细胞7～8d左右即可长满,并可稳定传代,传代细胞5～6d即可传代。诱导培养细胞碱性磷酸酶染色,骨连接素、骨桥素免疫细胞化学染色阳性。结论兔骨髓间充质干细胞经合理的体外诱导培养后,符合成骨细胞的形态特征和生物学特性,可为体外构建组织工程化骨提供自体种子细胞。

Experimental study of the inducing BMSCs to be osteoblasts

Objective To observe the growth and osteogenic property of cultured rabbit bone mesenchymal stem cells (BMSCs). Methods The bone marrow were obtained by aspirating from a healthy male rabbit, mononuclear cells were then isolated by Percoll density gradient centrifugation. The adhesive cells were preserved for passage. After passaging stably, the BMSCs were subcultured in a condition medium including dexamethasone, vitamin C and β-glycerophosphate. An inverted microscope, immunocytochemical staining for alkaline phosphatase (ALP) and immunocytochemical staining for osteonectin and osteopontin expression were applied to measure the BMSCs proliferation capability and osteogenic property. Results The BMSCs reached confluence and started to form multilayers on the 7th or 8th day, passaged stably as well. The BMSCs passaged within 5 or 6 days. The BMSCs in induced culture showed immunocytochemical staining for ALP and osteonectin and osteopontin expression. Conclusion The induced cultured rabbit BMSCs have the morphological and biological property of osteoblasts, which provides autogenous seeding cells for in vitro construction of tissue engineered bone.