目的 观察并鉴定兔骨髓间充质干细胞生物特性,建立一套简单、高效的培养方法。方法 采用直接贴壁法分离培养兔骨髓间充质干细胞,倒置显微镜下观察其形态。MTT法测定其增殖水平并绘制其生长曲线。流式细胞仪测定第三代骨髓间充质干细胞表型。结果 细胞培养第5 d,镜下见细胞呈小集落生长,形态呈多边形;7~8 d小集落融合成大集落,10 d左右可传代。第二代细胞镜下呈梭形成纤维样细胞,即间充质干细胞;细胞生长曲线成S形;流式细胞仪检测细胞表达CD 73、CD 90、CD 105、不表达CD 34、CD 45。结论 改良直接贴壁法可获得大量、纯化、稳定的间充质干细胞。

Objective To observe and identify the biological characteristics of rabbit bone marrow mesenchymal stem cells, and establish a set of simple , viable culture methods. Methods Rabbit bone marrow mesenchymal stem cells were extracted through adherent method directly, and use the inverted microscope to observe period morphology. The value-added level of the cells was assayed by MTT and the growth curve was drawn.The third passage of bone marrow mesenchymal stem cell phenotypes were measured by flow cytometry. Results Under the microscope,cells were small colony growth with shape of polygons when cells were cultured at the 5th day.Small colony confluent colony at 7-8 th day.The cells could be passaged at about 10th day.The second passage of cells showed long fusiform and fibroblast-like growth,that means marrow mesenchymal stem.The cell growth curve was S-shaped.Flow cytometry Results showed that the cells expressed CD 73,CD 90,CD 105, and did not express CD 34,CD 45. Conclusions The directly adherent method can get apundant, purified, and stable mesenchymal stem cells .