内皮细胞特异性缺失PTEN基因对小鼠胚胎AGM区血液血管干细胞发育的影响  

Effect of Endothelium-specific Deletion of PTEN on Hemangioblast Development in Mouse Embryo AGM Region

作　　者：高娇[1] 要晖宇[2] 梁晓雷[2] 王晓燕[2] 吴英[2] 刘元林[2] 毛宁[2] 

机构地区：[1]解放军第306医院检验科,北京100101 [2]军事医学科学院基础医学研究所细胞生物学研究室,北京100850

出　　处：《中国实验血液学杂志》2011年第5期1230-1233,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(编号30800427,30730043,81070393)

摘　　要：本研究探讨内皮细胞特异性缺失PTEN基因能否影响小鼠胚胎AGM区血液血管干细胞(hemangioblast)的发育。基于Cre/loxP系统,获得Tie2CrePtenloxp/loxp和Tie2CrePtenloxp/wt基因型小鼠胚胎。采用PCR方法进行基因型鉴定;分离E10-E10.5胚胎AGM区,用胶原酶消化成单细胞,在甲基纤维素半固体体系里进行BL-CFC(blastcolony-forming cell)培养,4-5天后计数BL-CFC数目。将单个BL-CFC挑出进行扩增培养,悬浮细胞进行造血集落培养,贴壁细胞在Matrigel上进行体外成血管实验,鉴定其双向分化功能。结果表明:内皮细胞特异性缺失PTEN基因,小鼠胚胎AGM区BL-CFC数量下降;同时,BL-CFC的造血分化能力增强,但BL-CFC来源的内皮细胞体外成血管能力下降。结论:内皮细胞特异性缺失PTEN基因,可以在血液血管干细胞水平影响造血和内皮细胞的分化。This study was aimed to investigate whether endothelium-specific deletion of PTEN can affect hemangioblast development in the AGM region of mouse embryos.Based on Cre/loxP system,the Tie2CrePtenloxp/loxp and Tie2CrePtenloxp/wt mouse embryos were obtained.The genotype was identified by PCR.After treated with type Ⅰ collagenase,the AGM region was dispersed into single-cell suspension,and then was cultured in blast colony-forming cell（BL-CFC） media.The number of BL-CFC was counted 4 or 5 days later.The hematopoietic capacity of BL-CFC was detected in methylcellulose culture system and the endothelial potential was assessed by tube-like structure formation on Matrigel.The results showed that the number of BL-CFC in AGM region of Tie2CrePtenloxp/loxp mouse embryo decreased as compared with Tie2CrePtenloxp/wt embryo.Whereas the hematopoietic capacity of mutant BL-CFC was enhanced,the endothelial potential,as evaluated by tube-like structure formation in vitro,was significantly reduced.It is concluded that the endothelial PTEN is capable of exerting regulatory functions on both the numbers and the dual potential of hemangioblast in mouse AGM region.

关 键 词：血液血管干细胞 AGM区 BL-CFC 

分 类 号：R331.2[医药卫生—人体生理学]