家兔骨髓基质干细胞离心法分离培养和鉴定  

Study on isolation and cultivation of rabbit bone marrow mesenchymal stem cells through gradient centrifugation

作　　者：马钊[1] 胡向东[2] 费琦[1] 张效栋[1] 张海燕[3] 孟海[1] 王奇[1] 杨雍[1] 王炳强[1] 

机构地区：[1]首都医科大学附属北京友谊医院骨科,北京100050 [2]首都医科大学附属北京友谊医院超声科,北京100050 [3]首都医科大学细胞生物学系,北京100069

出　　处：《临床和实验医学杂志》2015年第16期1330-1332,共3页Journal of Clinical and Experimental Medicine

基　　金：2013北京市优秀人才培养资助(项目号:2013D003034000005)

摘　　要：目的对家兔骨髓间充质干细胞进行分离培养的研究,并对得到的兔骨髓间充质干细胞进行鉴定。方法采用分层离心法分离原代间充质干细胞,经反复传代进行纯化,以免疫组化荧光染色法鉴定得到的细胞。结果通过贴壁法培养得到的兔骨髓细胞较为均一,可在体外进行长期培养传代,免疫组化显示细胞表达CD105、CD44,而不表达CD34。结论单纯分层离心提取并经过贴壁培养可得到较均一的兔骨髓间充质干细胞,可用于进一步的成骨等方面的研究。Objective To establish the method for isolation and cultivation of rabbit bone marrow mesenchymal stem cells（ BMSCs）.Methods The samples of bone marrow tissue were taken from rabbits,and BMSCs were isolated by using density gradient centrifugation. Purified BMSCs could be got through repeated subculturing. Immunohistochemical staining had been applied to observe cell surface antigens CD105,CD44 and CD34. Results The cultured cells had adherent growth ability with the morphology of fibroblast- like cells and spindle- shaped appearance.The positive expression of CD105 and CD44 was detectable while CD34 was not detectable. Conclusion The cultured cells are uniformly and purely undifferentiated BMSC which can be used in further study such as bone formation etc.

关 键 词：家兔 骨髓间充质干细胞 分离 培养 

分 类 号：R68[医药卫生—骨科学]