大鼠骨髓间充质干细胞的生物学特性研究

目的 建立大鼠骨髓间充质干细胞(MSCs)分离和培养扩增的方法。研究MSCs的生物学特性，为利用MSCs作为种子细胞治疗多种疾病提供实验基础。方法通过密度梯度离心、贴壁筛选法分离培养大鼠MSCs，测定其接种贴壁率；在倒置显微镜下连续观察细胞的形态变化，利用MTT法测定MSCs生长曲线；并应用流式细胞术测定细胞周期和表面标记物。结果MSCs体外培养生长状况良好，具有活跃的增殖能力，接种10h后贴壁率为96％以上；细胞周期显示有91．4％处于G0／Gl期；培养的MSCs阳性表达CD29、CD44，阴性表达CD34、CD45。结论在实验条件下，体外培养8代以前的MSCs生长稳定，增殖较快，细胞周期表现出较早期细胞特点，可作为组织工程的种子细胞。

The biological characteristics of rat bone marrow-derived mesenchymal stem cell.

Objective To establish a method for isolation and cultivation of mesenchymal stem cells from rat in vitro and explore their biological properties in order to provide an experimental basis for applying MSCs to achieve the therapeutic benefit as seed cells in some diseases. Methods MSCs were obtained from tibias and temurs by density gradient centrifugation and were tested the rate of adhesion. The configuration was observed under phase-contrast microscope consecutively and the cell growth was measured by MTT method. Cell cycle and surface antigens were detected through flow cytometer. Results The subcultured cells showed active proliferative ability. More than 96% of subcultured MSCs were adhesive in 10h. The cell cycle analysis showed that 91.4% of MSCs was in G_0/G_1 phase. MSCs were positive for CD29?CD44 and negative for CD11b?CD45. Conclusion In present culture condition, MSCs showed stable and rapid growth in vitro before 8th generation and the cell cycle also exhibit the infantility for MSCs. All the characteristics make it possible to MSCs to be the seed cells for tissue engineering.