小鼠胚胎干细胞来源的HPP-CFC体外和体内造血活性分析  

The Investigation of Hematopoietic Capacity of HPP-CFC Derived from Murine Embryonic Stem Cells in vitro and in vivo

作　　者：刘兵[1,2] 侯春梅[1] 吴英[1] 张双喜[1] 毛宁[1] 

机构地区：[1]军事医学科学院基础医学研究所细胞生物学研究室 [2]第二军医大学基础部免疫学教研室上海200433

出　　处：《生物工程学报》2003年第3期312-316,T001,共6页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划"863"资助项目 (No .2 0 0 1AA2 1713 1);国家重点基础研究发展规划"973"资助项目 (No .G19990 5 43 0 2 )~~

摘　　要：小鼠的造血系统起源于胚胎发育 7d的卵黄囊胚外中胚层 ,研究表明胚胎干细胞 (Embryonicstemcells ,EScells)体外分化模型能够模拟卵黄囊造血的发生过程 ;此外 ,诱导ES细胞体外定向造血细胞分化对于建立治疗性克隆以治愈多种血液病具有重要的研究和应用价值。高增殖潜能集落形成细胞 (Highproliferativepotentialcolony form ingcells ,HPP CFC)是体外培养的最原始的多潜能造血前体细胞之一。本研究发现 :小鼠ES细胞在体外分化 5～14d形成的拟胚体中含有HPP CFC。其再生潜能与胚胎期 9d的卵黄囊来源的HPP CFC相似 ,与骨髓来源则不同。RT PCR分析表明 :ES细胞来源的HPP CFC表达与造血干细胞增殖相关的特异性转录因子和多种造血生长因子受体。但分化 12d的拟胚体细胞和HPP CFC集落细胞移植受致死剂量照射的小鼠不能产生典型的脾结节。因此 ,ES细胞来源的HPP CFC在体外和体内造血活性的差异值得更深入地研究。The hematopoietic system of the mouse arises from extraembryonic mesoderm that migrate through primitive streak to the presumptive yolk sac at day 7 0 of gestation. However, the mechanisms regulating mesoderm commitment to hematopoietic lineages remain poorly understood. Previous studies demonstrated that the development kinetics and growth factor responsiveness of hematopoietic precursors derived from embryonic stem cells (ES cells) is similar to that found in the yolk sac, indicating that the onset of hematopoiesis within the embryoid bodies (EBs) parallels that found in the embryo. Furthermore, in vitro differentiation of ES cells to hematopoietic cells is valuable for establishment of therapeutic clone against a variety of hematological disorders. Despite the identification of multipotential hematopoietic progenitors in EBs, a subset of more primitive progenitors, identical to the high proliferative potential colony forming cells (HPP CFC) derived from human and murine hematopoietic tissues, have not been clearly identified regarding particular their replating potential in vitro. HPP CFC is among the most primitive hematopoietic multipotent precursors cultured in vitro. In this study, our aim was to investigate the in vitro and in vivo hematopoietic capacity of HPP CFC within the day 12 EBs, rather than the expansion of more committed progenitors. In this study the HPP CFC could be detected within EBs differentiated for 5 to 14 days of murine ES cells, but the development dynamics of the HPP CFC differed greatly among distinct serum lots. Qualitatively HPP CFC is capable of forming secondary colonies. As to our expectation the ES cells derived HPP CFC demonstrated similar regeneration capacity to those from yolk sac, giving rise to secondary granulocyte, erythrocyte, macrophage and mast cells, however largely differed from the counterparts of adult bone marrow. In addition, by RT PCR ES cells derived HPP CFC were found to express transcription factors associated closely with stem cell pro

关 键 词：胚胎干细胞 高增殖潜能集落形成细胞 脾结节形成单位 小鼠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]