【摘要】 目的观察小鼠骨髓内皮细胞条件培养液（ｍＢＭＥＣ－ＣＭ）对卵黄囊造血干／祖细胞体外生长的支持作用。方法取妊娠第８．５天的卵黄囊，经消化后获得卵黄囊细胞，取卵黄囊非贴壁细胞加入含１０％ｍＢＭＥＣ－ＣＭ和１０％ＦＢＳ的ＤＭＥＭ中培养２４ｈ后，收集细胞做半固体培养。结果ｍＢＭＥＣ－ＣＭ在液体培养体系中能扩增卵黄囊造血干／祖细胞使粒－巨噬系集落形成单位（ｇｒａｎｕｌｏｃｙｔｅ－ｍａｃｒｏｐｈａｇｅ　ｃｏｌｏｎｙ　ｆｏｒｍｉｎｇ　ｕｎｉｔ，ＣＦＵ－ＧＭ）和高增殖潜能集落形成细胞（ｈｉｇｈ　ｐｒｏｌｉｆｅｒａｔｉｖｅ　ｐｏｔｅｎｔｉａｌ－ｃｏｌｏｎｙ　ｆｏｒｍｉｎｇ　ｃｅｌｌ，ＨＰＰ－ＣＦＣ）的产率增加，经２４ｈ液体培养后的ＣＦＵ－ＧＭ和ＨＰＰ－ＣＦＣ的数目分别为培养０ｈ对照组的１１９．５％和１３０．８％（Ｐ＜０．０５＝；ｍＢＭＥＣ－ＣＭ联合ｆｌｔ３配基（ｆｌｔ３ｌｉｇａｎｄ，ＦＬ）和血小板生成素（ｔｈｒｏｍｂｏｐｏｉｅｔｉｎ，ＴＰＯ）后，其扩增ＣＦＵ－ＧＭ和ＨＰＰ－ＣＦＣ的能力明显增强，ｍＢＭＥＣ－ＣＭ联合ＦＬ和ＴＰＯ经２４ｈ液体培养后的ＣＦＵ－ＧＭ和ＨＰＰ－ＣＦＣ的数目分别为培养０ｈ对照组的１３２．０％和１７６．９％?更多还原

【Abstract】 Objective To investigate the effects of murine bone marrow endothelial cell conditioned me dium(mBMEC-CM)on the growth of yolk sac hematopoietic progenitors.Methods The serum-free mBMEC-CM was obtained from subcultures of murine endothelial cell line derived from bone marrow which was established in our laboratory.The murine yolk sacs were harvested on day8.5postcoitus(p c)and incubated with0.1%collagenase in10%fetal calf serum at37℃for40minutes.Yolk sac cells were incubated in tissue culture dishes at37℃for1hour.Nonadherent cells were collected for sem isolid culture assay of granulocyte-macrophage colony forming unit(CFU-GM)and high proliferative potential-colony forming cell(HPP-CFC)after being cultured in DMEM with10%mBMEC-CM and10%FBS for24hours.The number of CFU-GM and HPP-CFC was counted at day7and day14respecti vely.Results The growth of CFU-GM and HPP-CFC was supported by mBMEC-CM with GM-CSF.mBMEC-CM could induce theproliferation and differentiation of yolk sac hematopoietic stem cells and progenitors in liquid culture system.The percentages of CFU-GM and HPP-CFC compared with the0hour control were(119.5±5.7)%and(130.8±9.8)%respectively after24hours liquid culture (P<0.05). The expansion effects of mBMEC-CM on CFU-GM and HPP-CFC were enhanced by compounded with flt3ligand(FL)and thrombopoietin (TPO).The percentages of CFU-GM and HPP-CFC compared with the0hour control were (132.0±6.2)% and (176.9±12.8)% respectively after24hours liquid culture(P<0.01=.Conclusion Murine bone marrow endothelial cell conditio-ned medium could support the growth and proliferation of yolk sac hematopoitic stem cells and progenitors,and this promoting effect was further enhanced by addition of FL and TPO.更多还原