细胞因子介导的体外扩增对脐血干/祖细胞粘附分子表达的影响  

The Effects of Cytokines Mediated ex vivo Expansion on the Cell Adhesion Molecule Expression of Cord Blood Hematopoietic Stem and Progenitor Cells

作　　者：翟琼莉[1] 邱录贵[1] 刘燕[1] 李茜[1] 韩俊领[1] 周征[1] 李新[1] 应红光[1] 韩忠朝[1] 

机构地区：[1]中国医学科学院中国协和医科大学血液学研究所实验血液学国家重点实验室,天津300020

出　　处：《中国医学科学院学报》2002年第1期7-10,共4页Acta Academiae Medicinae Sinicae

基　　金：攀登计划(95-专-10);天津市科技发展计划项目(003119811)~~

摘　　要：目的比较脐血（ＣＢ）ＡＣ１３３＋细胞扩增前后ＣＤ３４＋　细胞亚群表面归巢相关粘附分子ＶＬＡ－４（ＣＤ４９ｄ）、ＶＬＡ－５（ＣＤ４９ｅ）、ＬＦＡ－１（ＣＤ１１ａ）、Ｌ－ｓｅｌｅｃｔｉｎ（ＣＤ６２Ｌ）和ＰＥＣＡＭ－１（ＣＤ３１）等的表达情况，以评价细胞因子介导的体外扩增对干／祖细胞（ＨＳＰＣ）归巢功能的影响。方法将从新鲜ＣＢ标本中纯化的ＡＣ１３３＋　　　　细胞接种于无血清培养基ＱＢＳＦ－６０的无基质悬浮体系培养扩增１４ｄ，加入早期作用因子ＦＬ、ＳＣＦ和ＴＰＯ组合（ＦＳＴ），并在接种０ｄ时添加一剂ＩＬ－３，分别于培养０，７、１０和１４ｄ检测扩增潜能和上述几种粘附分子的表达情况。结果（１）在１４ｄ的培养扩增中，各阶段的ＨＳＰＣ均得到有效扩增，至１４ｄ时ＡＣ１３３＋和ＣＤ３４＋细胞分别增加３３．５０和６４．５６倍；（２）表达上述粘附分子的各ＣＤ３４＋　　　　细胞亚群均有不同程度（约２０～１６０倍）的扩增；（３）扩增后ＣＤ３４＋　细胞表面的粘附分子ＣＤ１１ａ、ＣＤ４９ｅ和ＣＤ４９ｄ的表达与原代ＣＤ３４＋细胞持平或上升，而ＣＤ６２Ｌ和ＣＤ３１的表达则有不同程度的下调。结论我们建立的短期培养体系不仅可以支持ＣＢ　ＨＳＰＣ的?Objective To compare the expression of cell adhesion molecules(CAMs)among VLA-4(CD49d),VLA-5(CD49e),LFA-1(CD11a),L-selectin(CD62L),and PECAM-1(CD31)which are more related to the homing of hematopoietic stem and progenitor cells(HSPC)on the ex vivo expan ded CD34 + subset with that of fresh isolated AC133 + cells.Methods AC133 + cells selected from fresh cord blood(CB)samples were cultured in QBSF-60serum-free media in the presence of Flt-3ligand+SCF+TPO(FST),with initial addition of IL-3for up to2week.Expansion potential and the expression of above CAMs were evaluated at day0,day7,day10and day14.Results(1)Sim ultaneously numerical expansion of various HSPC was constantly observed during the culture,and the fold expansion of AC133+ cells and CD34+ cells on day14were33.50 and 64.56respectively; (2)The number of CD34+ subsets expres-sing the above adhesions were all increased at different degrees(from20fold to160fold).(3)The expressions of CD11a,CD49d,and CD49e on ex vivo exp anded CD34+ cells were increased as compared to their baseline levels,but the percentage of CD62L+ and CD31+subpopulations in CD34+ cells were decreased.Conclusions Our short-term cul ture system can not merely support the simultaneous expansion of CB derived AC133+ cells,but the expanded hematopoietic progenitors may well sustained the expression of homing-related adhesion molecules.

关 键 词：脐血 AC133^+细胞 扩增潜能 粘附分子 干细胞/祖细胞 细胞因子介导 

分 类 号：R331.14[医药卫生—人体生理学]