全骨髓法培养C57小鼠骨髓间充质干细胞的生物学特性

背景：饲养层细胞多数利用丝裂霉素C来处理，虽丝裂霉素C抑制了细胞增殖，但是细胞仍处于有分泌功能的活性状态，能够分泌不同的细胞因子。而骨髓中的非间充质骨髓细胞和血浆中的各种分泌因子，保持间充质细胞生长的微环境，提高间充质干细胞的产量。 目的：探究全骨髓培养法分离培养的小鼠骨髓间充质干细胞的生物学特性。 方法：采用全骨髓贴壁法纯化扩增C57小鼠的骨髓间充质干细胞，检测细胞增殖的动力学变化、细胞表面的免疫标志物、多向分化的潜能及细胞周期的变化。 结果与结论：全骨髓培养法获得的小鼠骨髓间充质干细胞具有黏附于塑料培养器皿的能力，流式细胞仪检测表达CD45，CD105和Sca-1，不表达CD34，CD133和C-kit等间充质干细胞的表面标记特征；细胞倍增时间（57.11＆#177;1.5） h；诱导后具有向成骨细胞、脂肪细胞和软骨细胞分化的能力；细胞周期分析表明64%的细胞处于G 0-G 1期。提示全骨髓培养法获取的C57小鼠骨髓间充质细胞具有间充质干细胞的生物学特征。

Biological characterization of C57 mouse bone marrow mesenchymal stem cells using a whole bone marrow adherent culture technique

BACKGROUND：Under mitomycin C treatment, feeder cells appear to have restricted proliferation, but they are stil able to secret different cytokines. Non-mesenchymal stem cells from the bone marrow and secreted factors in plasma maintain the micro-environment suitable for the growth of mesenchymal stem cells that can improve the yield of mesenchymal stem cells. OBJECTIVE：To study the biological characteristics of C57 mouse bone marrow mesenchymal stem cells isolated using a whole bone marrow adherent culture technique. METHODS：Using the whole bone marrow adherent culture technique, purified and amplified C57 mouse bone marrow mesenchymal stem cells were harvested. cellproliferation kinetics, immune cellsurface markers, multiple differentiation potential and cellcycle were detected. RESULTS AND CONCLUSION：Using the whole bone marrow culture, mouse bone marrow mesenchymal stem cells were harvested and capable of adhering to the plastic culture vessel. The obtained cells expressed CD45, CD105 and Sca-1, but were negative for CD34, CD33 and C-kit. The doubling time was （57.11＆#177;1.5） hours. The cells could be induced to differentiate into osteoblasts, adipocytes and chondrocytes. The cellcycle analysis showed that 64%of cells were in G 0-G 1 phase. These indicates that C57 mouse bone marrow mesenchymal stem cells isolated using a whole bone marrow adherent culture technique have biological characteristics of mesenchymal stem cells.