摘要
目的比较不同方法获取骨髓间充质干细胞的效率,为其在组织工程的应用确立最佳培养方案。方法以3个月龄新西兰大白兔为实验对象,通过对全血培养法、溶血纯化法、密度梯度离心法进行比较性研究,对克隆形成率、首次传代时间、扩增成功率等指标进行比较。结果对克隆形成率和扩增成功率而言,溶血纯化方法最低传代时间为20.5d,全血培养方法有部分提高传代时间为(13.9±2.9)d;而密度梯度离心方法的效率最高,首次传代时间为(7.5±0.7)d。结论对于成年动物穿刺获取骨髓间充质干细胞而言,密度离心>全血培养>溶血纯化方法,明确了一种实用有效的骨髓间充质干细胞分离培养方法。
Aim To compare the efficiency of different methods in culturing mesenchymal stem cells(MSCs)derived from bone marrow as a basis of the optimal isolation method of tissue engineering.Methods 12 Zelanian rabbits, aged 3 months, were randomly divided into 3 groups of 4 subjects: group A,group B and group C.The whole blood culture, hemolysis and density gradient centrifugation protocols were used to culture mesenchymal stem cells among 3 groups respectively. The cloning formation rate, primary passage time and success rate of each method were observed.Results For the cloning formation rate and primary passage time, the best time of passage given by the hemolysis method was 20.5 d, partial rising time of passage given by the whole blood culture method was (13.9±2.9) d, the most efficient time of passage provided by density gradient centrifugation method was (7.5±0.7) d.Conclusion The gradient centrifugation protocol was the most preferable method for the isolation of mesenchymal stem cells derived from bone marrow in adult rabbits, and this study provides a reliable method for the adult MSCs culture.
出处
《中国临床康复》
CSCD
2003年第11期1636-1637,共2页
Chinese Journal of Clinical Rehabilitation
