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PAGE PAGE 8 兔骨髓间充质干细胞分离、体外培养及其生物学活性的观察Isolate, Culture Rabbit Bone Marrow Mesenchymal Stem Cells And Observe Biological Activity In Virtue.摘要目的：研究兔骨髓间充质干细胞（Rabbit bone marrow mesenchymal stem cell，rBMSCs）的体外分离，培养及其生长活性的观察，为实验提供种子细胞。。方法：戊巴比妥耳缘静脉麻醉下，在无菌条件下用12号骨穿针在股股大转子处抽取骨髓液，液密度梯度离心法+全骨髓贴壁培养法相结合的方法进行体外培养及传代。分别取第1代和第4代细胞做MTT检测和流失细胞仪检测。结果：接种48小时后观察，见少量细胞呈长梭性，大部分为短梭形和多边形。接种72小时观察，贴壁细胞明显曾多，呈长梭形。接种第5天，细胞增殖明显，并且可见若干细胞团落星成。接种第7天，达到融合。第1，4代细胞经MTT和流式细胞仪检测，两组的增殖能力和生长周期无显著性差异（P0.05）结论：percoll分离液+全骨髓贴壁培养法可以成功的分离出骨髓rBMSCs，，其在体外培养具有活跃的增殖能力。传代后细胞的生物学形态基本一致。关键词：干细胞，分离，培养，生物学活性。Abstract: To research a method to isolate, culture bone marrow mesenchymal stem cells. Observe biological activity of bone marrow mesenchymal stem cells in vitro. To provide seed cells for the experiment. Method: The rat was narcotized by pentobarbital injected into the vessel of ear. Under the sterile condition, the 12 modullo-puncture needle was used to extract marrow fluids at the great trochanter. Bone marrow mesenchymal stem cells were isolate and culture by density gradient centrifugation and plastic method. The proliferation ability and growth cycle of P1 and P4 cells was analyzed by MTT and flow cytometry.Result: After seed 48h later , some cells were long spindle, most cells were spindle and small polygonal, 72h later, plastic cells increase obvious, the cells showed long spindle, 5 days later ,the cells grew obvious and formed some cell group,7 days later, the cells got confluence. The P1 and P4 cells were tested by MTT and flow cytometry, the result showed that the 2 groups had no significant different (P0.05) in proliferation ability and growth cycle.Conclusive：Density gradient centrifugation and plastic method can isolate and culture bone marrow mesenchymal stem cells successfully. The cells can proliferate actively in vitro. After passage, the biological character of the cells had no change.Key words: stem cells，isolate，culture，biological activity。前言骨髓间充质干细胞（bone marrow mesenchymal stem
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