骨髓间充质干细胞减轻体外扩增的脐血造血干细胞衰老的作用  

The protection of senescence by bone marrow mesenchymal stem cells in human cord blood CD34＋ expanded cells

作　　者：王彩霞[1] 张玉平[1] 陈小卫[1] 杜庆华[1] 潘世毅 黎宇苗[1] 应逸[1] 王顺清[1] 毛平[1] 

机构地区：[1]广州医科大学附属市一人民医院血液内科,510180

出　　处：《中华器官移植杂志》2013年第7期432-435,共4页Chinese Journal of Organ Transplantation

基　　金：广州市医药卫生科技项目（201102A213087）

摘　　要：目的探讨骨髓间充质干细胞（MSC）对脐血造血干细胞体外扩增及抗细胞衰老的作用。方法从新鲜脐血中分离出CD34＋细胞，分别接种在含或不含MSC及细胞因子的培养体系中，取培养10d后的造血干细胞，分别用于细胞计数，集落分析，流式细胞仪检测表面标记，并对培养后的细胞进行p半乳糖苷酶活性检测，采用实时定量聚合酶链反应（PCR）检测衰老相关基因p16INK4amRNA的表达。结果体外培养10d后，MSC组、细胞因子组及MSC＋细胞因子组对脐血总有核细胞（TNC）、CD34＋细胞的体外扩增及克隆形成能力均有支持作用，以MSC十细胞因子组最为明显（P〈0．05），其扩增倍数分别达到（45．3±6．8）倍、（38．4±5．8）倍及（50．2±4．2）倍，但衰老细胞比例及p16INK4smRNA表达倍数则以MSC组最低，MS（2＋细胞因子组次之，细胞因子组两项指标均为最高（P〈（）．05）。结论MS（2能更好保护体外扩增的脐血造血干细胞，减少细胞衰老的发生，Msc＋细胞因子是有效扩增脐血造血干细胞并保持细胞干性的更理想方法。Objective To investigate the effects and mechanisms of bone marrow mesenchymal stem cells （MSCs） on cord blood hematopoietic cells after culture ex vivo with or without cytokines, and and develop an optimal culture system for cord blood hematopoietic cells culture ex vivo. Method CD34＋ cells separated from cord blood were cultured for 10 days. Total nucleated cells （TNCs）, CD34＋ cells and colony-forming unit （CFU） were analyzed. SA-13-gal and p16 mRNA was detected by using real-time reverse transcriptase （PCR）. Results Ex vivo results demonstrated that the three culture systems supported expansion of TNCs, CD34＋ cells and CFU, especially in the MSCs ＋ cytokine group. The proportion of senescent cells and pl 6 mRNA expression in the expanded cells in the MSCs group were lower than those in the other two groups （P〈0. （15）. Conclusion MSCs are capable of protecting hematopoietic cells and reduce the occurrence of cell senescence in expanded cells. MSCs in combination with cytokines could efficiently support proliferation of cord blood hematopoietic cells cultured ex vivo.

关 键 词：脐血干细胞移植 细胞培养技术 细胞衰老 间质干细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]