人脐血间充质干细胞鉴定及用5氮胞苷诱导后的生物学特性研究.docVIP

人脐血间充质干细胞鉴定及用5-氮胞苷诱导后的生物学特性研究 　　作者：黄景玲 作者单位：青海省心血管病专科医院 　　【摘要】 目的 探讨脐血间充质干细胞经体外扩增纯化表面CD标志的鉴定及在体外定向分化成心肌样细胞的诱导剂5-氮胞苷(5-aza)作用后的生物学特性。方法 健康妊娠产妇分娩的胎儿脐带血，分离单个核细胞进行培养、传代，取第3代以后细胞用流式细胞仪直接标记分析CD34、CD44、CD90细胞表面标志。10Μmmol/L5-aza诱导4周后，进行透视电镜观察，并检测细胞肌球蛋白重链基因的表达。结果 体外纯化扩增后的脐血间充质干细胞含有与骨髓来源的间充质干细胞相同的细胞表面标志，第3代以后细胞纯度较高，经5-aza体外诱导培养可形成心肌样细胞。结论 脐血间充质干细胞经体外纯化扩增，第3代以后的细胞纯度较高，在体外经5-aza诱导定向分化为心肌样细胞后，其基本的生物学特性未发生改变。 　　【关键词】 脐血间充质干细胞 5-氮胞苷 生物学特性 心肌样细胞 　　APPRAISAL OF HUMAN MSCS FROM UCB AND ITS 　　BASIC BIOLOGIC CHARACTERIZATION AFTER 　　5-AZACYTIDINE TREATMENT 　　Huang Jingling 　　(Qinghai Cardiovascular Hospital) 　　Abstract Objective To Detect the human MSCs markers of CD34、CD44、CD90 in Cultured and expansived stem cells (MSCs) from umbilical cord blood (UCB) in vitro and maintain its biological characterization during expansion and to induce cardiomyogenic differentiation. Methods We have isolated single cell-derived, after the thirth-passage cells were harvested, and labeled with antibodies against human antigens CD34, CD44, CD90 as indicated and analyzed by FACS. The morphology changed under transmission electron micrograph well-organized myofilaments ties, and sarcomeres after 5-azacytidine treatment. Four weeks inducing, these cells expressed the myocardial marker of myosin heavy chain. The growth curve drawing and Flow cytometry for cell cycle analysis both UCB-derived MSCs and UCB-derived MSCs after 5-azacytidine treatment. Results After the thirth-passage UCB -derived cells is MSCs, the cells maintain their stem cell nature during expansion and differentiation into in vitro. The immunophenotype of these clonally expanded cells is consistent with that reported for bone marrow mesenchymal stem cells. Conclusion Umbilical cord blood can serve as a valuable and convenient source of cariomyocyte mesenchymal stem cells. 　　Key word UCB-derived MSCs 5-aza differentiation Biologic Bharacterization 　　干细胞体外诱导定向分化为心肌细胞的试验中，常用的诱导介质有心肌细胞的提取物、5-氮胞苷(5-aza)、二