造血发育过程中造血祖细胞生物学特性的时空变化  

Biological characteristics of hematopoietic progenitor cells at different stages of hematopoietic development

作　　者：周琨[1] 黄丽芳[1] 胡采红[1] 刘文励[1] 孙汉英[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院血液科,武汉430030

出　　处：《生理学报》2009年第5期493-498,共6页Acta Physiologica Sinica

基　　金：supported by the National Natural Science Foundation of China(No.30570773);Doctoral Fund of Ministry of Education of China(No.20060487061;200804870008)

摘　　要：本实验以小鼠胚胎主动脉-性腺-中肾(aorta-gonad-mesonephros,AGM)区、胎肝(fetalliver,FL)以及小鼠骨髓(bonemarrow,BM)为研究对象,研究各阶段造血祖细胞在发育过程中的c-kit的表达变化和部位迁移,以及不同发育阶段、不同造血部位来源c-kit+细胞的生物学特性。用流式细胞术检测小鼠胚胎10.5天(embryonic10.5,E10.5)、E11.5的AGM区,E12.5、E14.5、E16.5、E18的FL以及BM来源的单个核细胞中c-kit+细胞的比例。用磁珠分选法分离E10.5AGM区、E14.5FL以及BM来源c-kit+细胞,贴壁培养法分离小鼠E14.5FL基质细胞。建立体外Transwell共培养体系,比较不同来源的c-kit+细胞增殖、分化、迁移以及集落形成能力。流式细胞术检测发现:相对于BM,E10.5AGM区和E12.5FL中c-kit+细胞比例较高。进行体外培养,AGM区来源c-kit+细胞具有较强的增殖潜能,而FL、BM来源的c-kit+细胞在体外培养中更容易分化;AGM区和FL来源的c-kit+细胞比BM来源的c-kit+细胞在体外培养中具有更强的趋化迁移能力。集落形成能力方面,AGM区来源c-kit+细胞具有较强形成混合集落形成单位(CFU-Mix)的能力;FL、BM来源c-kit+细胞则能形成更多粒单系集落形成单位(CFU-GM)。以上结果表明,和FL、BM来源的c-kit+细胞相比,AGM区来源c-kit+细胞具有更强的体外增殖、多系分化潜能以及趋化迁移能力,在未来可能有着更广阔的应用前景。The aim of the present paper is to better understand the mechanism of hematopoietic development through.studying the biological characteristics of hematopoietic progenitor cells at different stages of development. Firstly, the c-kit expression levels of the mononuclear cells from murine embryonic aorta-gonad-mesonephros （AGM） region at embryonic day （E）10.5 and E11.5, fetal liver （FL） at E12.5, E14.5, E16.5, E18 and bone marrow （BM） were assayed with fluorescence activated cell sorting （FACS）. Secondly, hematopoietic progenitor cells derived from AGM at E10.5, FL at E14.5 and BM were isolated by using c-kit microbeads. Isolated c-kit population cells from AGM, FL and BM were then co-cultured with E14.5 FL-derived stromal cells in transwell co-culture system in vitro. After 3, 7, 10 days of co-culture, numerous floating cells were generated. The floating cells generated in transwell inserts were collected for FACS cell count, migration activity detection and colony forming unit （CFU） formation assay. The results showed that the c-kit was highly expressed in El0.5 AGM, with the percentage of c-kit＋ cells declining during AGM development, c-kit expression was highly expressed again in E12.5 FL, declining along with the progressive development of the FL region. Co-cultured with FL-derived stromal cells, El0.5 AGM-derived c-kit＊ cells produced the highest number of hematopoietic cells, while BM-derived c-kit＋ cells produced the lowest number of hematopoietic cells. Compared with El0.5 AGM-derived c-kit＋ cells, E14.5 FL- and BM- derived c-kit＊ cells inclined to differentiate after 7 to 10 days of culture in vitro. E10.5 AGM and E14.5 FL-derived c-kit＋ cells exhibited a higher migration activity than BM-derived c-kit＋ cells. Moreover, E10.5 AGM-derived c-kit＋ cells showed a higher ability to form mixed colony-forming unit （CFU-Mix） colony. In conclusion, compared with FL- and BM-derived c-kit＋ cells, E10.5 AGM-derived c- kit＋ hematopoietic progenitor cells exhibit better p

关 键 词：造血祖细胞 造血发育 主动脉-性腺-中肾区 胎肝 骨髓 

分 类 号：R331.1[医药卫生—人体生理学] R329.28[医药卫生—基础医学]