应用免疫磁珠分离成人骨髓中STRO-1阳性细胞  

Isolation of STRO-1 positive cells in adult marrow by immunomagnetic selection

作　　者：李静[1] 章庆国[1] 郭宗科[1] 

机构地区：[1]东南大学临床医学院整形外科,江苏省南京市210009

出　　处：《中国临床康复》2006年第29期16-19,i0001,共5页Chinese Journal of Clinical Rehabilitation

基　　金：江苏省高新技术研究项目资助(BG2001033);2003年东南大学医学研究基金面上项目(502-2003YJ25)~~

摘　　要：目的:利用免疫磁珠分离成人骨髓中STRO-1+阳性细胞,获得同质性骨髓基质干细胞。方法:实验于2004-06/2005-06在东南大学修复重建外科研究所实验室完成。采集正常自愿献髓者骨髓,淋巴细胞分离液体离心纯化分离后利用免疫磁珠分离出STRO-1+细胞,研究其生长曲线、体外扩增能力、集落形成能力、细胞表型、细胞周期、成骨能力。结果:①利用免疫磁珠可从成人骨髓中获取STRO-1+细胞。经体外培养发现STRO-1+细胞贴壁生长,相差显微镜下STRO-1+细胞呈成纤维细胞梭形外观。②生长曲线可分为细胞生长延滞期、对数生长期及稳定期,细胞倍增时间为57h。③STRO-1+细胞可以在体外扩增12周左右,形成的成纤维细胞集落数、处于细胞分裂期的细胞数、扩增倍数均低于传统分离方法得到的骨髓基质干细胞。④经流式细胞仪检测,STRO-1+细胞稳定地表达CD29,CD44,CD105,不表达造血细胞系的表面标记CD14,CD34,CD45,HLA-DR。⑤经成骨培养发现STRO-1+的骨髓基质干细胞具有很强的成骨分化能力。结论:利用免疫磁珠能够从成人骨髓中快速分离、纯化骨髓基质干细胞,且具有高效、灵敏、对骨髓基质干细胞活性影响小的优点。AIM： To isolate homogeneous bone marrow mesenchymal cells （BMSCs） from adult bone marrow by inununomagnetic selection of STBO-1^＋ cells. METHODS： The experiment was conducted in the Laboratory of Reparative and Reconstruction Surgery of Southeast University between June 2004 and June 2006. Bone marrow obtained from normal voluntcem was centrifuged and purified with lymphocyte isolation liquid, from which STRO-1^＋ cells were isolated by magnetic immunomagnetic beads to study the growth curve, in vitro amplification capacity, colony forming capacity, cell phenotype, cell cycle as well as ostecgenic capacity. RESULTS： ①STRO-1^＋ cells could be obtained from adult bone marrow by immunomagnetic bead selection. It could be found from in vitro culture that STRO-1^＋ cells grew into monolayer cells adhering to the bottom of culture flask, and the cultured cells primarily were long spindle-shaped under phase-contrast microscope. ②The growth curve of STRO-1^＋ cells could be divided into three stages： arrearage, logarithm, and stationary, and the multiplication time of STRO-1^＋ cells was 57 hours. ③STRO-1^＋ cells were capable of in vitro multipling for 12 weeks, and number of colony in fibroblast, cell numbers in cell division phase as well as the amplification multiple were lower than those in BMSCs gained by plastic adberence.④ Flow cytometry inspection showed that STRO-1^＋ cells could uniformly express CD29, CD44, CD105 without expressed CD14, CD34, CD45 and HLA-DR.⑤TRO-1^＋ cells had strong potential of ostcogenic differentiation. CONCLUSION： Irnmunomagnetic beads selection can be used to isolate and purify BMSCs from adult marrow, which is effective, sensitive with less influence on the activity of BMSCs.

关 键 词：骨髓 干细胞 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]