摘要
目的：探索体外分离培养及纯化SD大鼠骨髓间充质干细胞（mesenchymal stromal cells，MSCs）的方法。方法：采用贴壁分离和消化控制相结合的方法，分离纯化3～4周龄大鼠MSCs，免疫细胞化学法鉴定其活性和表型。结果：原代培养的MSCs呈圆形、梭形、多角形等，24h内即可见少量细胞体贴壁伸展，7～8d左右可达90％融合，纯化后的MSCs呈均匀一致的长梭形，24h内细胞贴壁率99％，传代周期为7d左右。免疫细胞化学检测显示CD34表达阴性，CD44表达阳性。结论：贴壁分离和消化控制相结合能有效分离纯化成年大鼠MSCs，细胞纯度较高，细胞活力强，生物学性状稳定，是目前一种比较理想的分离纯化方法。
Objective To investigate the more effective methods of isolation, culture of mesenchymal stem cell （MSCs）, and examine the phenotype of MSCs in lab of Shenzhen Institute of Integrated Medicine of Shenzhen Second People＇s Hospital. Methods MSCs were separated from 3-4 weeks rats bone marrow by plastic adherence methods and purified by controlling the time of digestion combined with adhesion separation, and proliferated in culture flasks. Immunocytochemical method identified the MSCs active and phenotype. Results Primary cultured MSCs were oval, spindle-shaped or polygonal, and adhered to plastic surface within 24 h and reached 90% confluence within 7-8 days. After purification and proliferation, they were uniformly long spindle-shaped and passaged every seven days. The adhesion rate within 24 hours was 99%. lmmunocytochemistry showed MSCs were positive for CD44, while negative for CD34. Conclusion A feasible method for the isolation, purification and proliferation of MSCs from rat bone marrow has been established. The cultured MSCs are active in high purity with stable biological character. The culture method combining adhesion separation and digestion control is simply performed in need of little condition, so it is one of the optimal methods for cell preparation and purification at present.
出处
《深圳中西医结合杂志》
2008年第6期337-339,共3页
Shenzhen Journal of Integrated Traditional Chinese and Western Medicine
基金
深圳市科技计划资助项目(200802028)
