人胎盘CD133^＋细胞体外扩增巨核系祖细胞的研究  

In vitro expansion of megakaryocyte progenitor cells from human placenta CD133^＋ cells

作　　者：王丽[1] 陈代雄[1] 方宁[1] 章涛[1] 刘祖林[1] 刘金伟[1] 万卫红[1] 

机构地区：[1]遵义医学院附属医院、贵州省细胞工程重点实验室,563003

出　　处：《中华血液学杂志》2008年第9期615-618,共4页Chinese Journal of Hematology

基　　金：贵州省科技计划发展项目（2051;2003JGY005）

摘　　要：目的探讨人胎盘组织CD133^＋（PT-CD133^＋）细胞体外扩增分化巨核系祖细胞（MPC）的能力。方法采用磁珠激活细胞分选系统（MACS）分选PT—CD133^＋细胞，接种于含TPO、IL-3和SCF的无血清液体培养体系中体外扩增MPC，于培养7、10和14d进行细胞计数，流式细胞术检测细胞扩增过程中CD133、CD34、CD41抗原表达的动态变化，并采用半固体培养法对不同扩增阶段的细胞进行巨核祖细胞集落（CFU-MK）培养。结果培养至7d时，PT—CD133^＋细胞扩增效果最佳，扩增了（13±2）倍，培养至14d，细胞总数扩增了160倍；随着扩增时间的延长，CD133、CD34表达逐渐下调，而CD41的表达逐渐增强，扩增后的巨核系细胞多呈幼稚状态，未见血小板形成；PT—CD133^＋细胞扩增前后均能形成CFU—MK，扩增10d的细胞形成的CFU—MK总数最多，扩增了（54±10）倍。结论人PT—CD133^＋细胞具有体外扩增分化为MPC的能力，培养10d的细胞形成的CFU—MK总数最多。Objective To study the expansion potential of megakaryocyte progenitor cells （MPC） from human placenta tissue CD133^＋ （PT-CD133^＋ ）cells. Methods PT-CD133^＋ cells were purified from mononuclear cells（MNC） by magnetic activated cell sorting（MACS） and seeded in serum-free liquid culture medium supplemented with thrombopoietin （TPO）, interleukin-3（IL-3）, and stem cell factor（SCF） to expand MPC. At day 7, 10 and 14, the total cell number was counted and the dynamic changes of CD133, CD34, and CD41 antigens expression during ex-vivo expansion were analyzed by flow cytometry （FCM）. PT-CD133^＋ cells at different expansion time were collected and cultured in collagen semisolid medium for colony forming units-megakaryocyte （CFU-MK） assay. Results PT-CD133^＋ cells could be optimally expanded at day 7 by 13 ± 2 fold increase in serum-free liquid culture systems and the total cell number was expanded by 160 fold at day 14. With the expansion time going on, the expression of CD133, CD34 decreased and that of CD41 increased. The expanded megakaryocytes were immature and no sign of platelet formation. Both PT- CD133^＋ cells before and after expansion could form CFU-MK, the total number of CFU-MK peaked at day 10 of expansion by 54 ± 10 fold increase. Conelusion Human PT-CD133^＋ cells have a high capacity of MPC expansion, 10 days culture could give rise to the maximum number of CFU-MK.

关 键 词：造血干细胞 抗原 CD133 巨核细胞 细胞培养 胎盘 人 

分 类 号：R686[医药卫生—骨科学]