不同分离方法及首次换液时间对兔骨髓间充质干细胞生物活性的影响  

Effect of different isolation methods and time for first medium exchange on biological characteristics of rabbit bone marrow mesenchymal stem cells

作　　者：陈晓佩[1] 于文浩[1] 王亚丹[1] 刘远哲[1] 张宁[1] 王新庄[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450000

出　　处：《中国兽医学报》2015年第8期1307-1311,共5页Chinese Journal of Veterinary Science

基　　金：国家"863"计划资助项目;河南省基础与前沿技术研究计划基金资助项目(092300410081)

摘　　要：探讨不同分离方法及首次换液时间对兔骨髓间充质干细胞（BMSCs）生物活性的影响。分别采用红细胞裂解法和全骨髓贴壁法分离纯化兔骨髓间充质干细胞,倒置显微镜观察细胞形态、台盼蓝染色检测细胞活性、记录首次传代时间、细胞免疫组化法鉴定CD90、CD44、CD34抗原表达;分别于培养12,24,48,72h首次换液,以后每2d换液1次,记录各组0～3代培养时间。结果显示,2种分离方法均能得到较均一的长梭形细胞,聚集成旋涡状均匀生长,CD90、CD44抗原表达阳性,CD34抗原表达阴性。其中红细胞裂解法所得细胞活性较全骨髓贴壁法高,首次传代时间短,差异显著（P〈0.05）;24h首次换液各代达传代所需时间与其他各组比较差异极显著（P〈0.01）,为最佳首次换液时间,可显著缩短细胞培养周期。这表明,采用红细胞裂解法,24h首次换液能够提高骨髓间充质干细胞的活性,缩短细胞培养周期。To explore the influence of different separation methods and time for first medium exchange on biological activity of rabbit bone marrow mesenchymal stem cells（BMSCs）.BMSCs were obtained by red blood cell lysis and whole bone marrow culture.The cell morphology of BMSCs was observed under inverted microscope,the cell viability was detected by trypan blue staining,the first time of passage was recorded,the CD90,CD44,CD34 antigen expression were detected by immunocytochemistry technique.Time for first medium exchange was at 12,24,48,72 hrespectively,then all groups were exchanged every 2d.The period of each passage was measured.The result showed that BMSCs could be isolated by the two methods.The adhere cells showed shuttle or gathered into a vortex-like uniform growth,and positive for CD90,CD44 antigen,negative for CD34.BMSCs isolated by the red blood cell lysis were significantly superior to the whole bone marrow culture by the cells viability and primary culture time（P〈0.05）;time needed for each generation in 24 hfor first time medium exchange significantly shortende compared with other groups（P〈0.01）.The result showed that the red blood cell lysis method,24 hfor the first time medium exchanges can improve the activity of bone marrow mesenchymal stem cells,shorten the cycle of cell culture.

关 键 词：兔 骨髓间充质干细胞 红细胞裂解法 全骨髓贴壁法 

分 类 号：S852.1[农业科学—基础兽医学]