人胎盘组织来源造血干/祖细胞及其特性(英文)  

Properties of hematopoietic stem/progenitor cells derived from human placenta tissues

作　　者：章涛[1] 方宁[1] 陈代雄[1] 刘祖林[1] 刘金伟[1] 万卫红[1] 祁莹[1] 肖建辉[1] 肖瑜[1] 

机构地区：[1]遵义医学院附属医院贵州省细胞工程重点实验室,贵州省遵义市563003

出　　处：《中国组织工程研究与临床康复》2008年第21期4172-4176,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：贵州省科技计划发展项目(2051;2003JGY005)~~

摘　　要：背景:近年来的研究表明,除已知的人骨髓、外周血和脐带血中存在造血干/祖细胞外,人胎盘组织中也有造血干/祖细胞存在。目前为止,还缺乏对人胎盘组织造血干/祖细胞的增殖分化特性及人胎盘组织淋巴细胞亚群组成和免疫原性等的深入研究。目的:探究人胎盘组织是否含有比脐带血更丰富的造血干/祖细胞,并对其造血祖细胞系增殖分化能力进行检测,同时对人胎盘组织淋巴细胞亚群组成及表型特征进行分析。设计、时间及地点:开放性实验,于2004-01/2006-12在贵州省细胞工程重点实验室完成。材料:经产妇知情同意,无菌采集遵义医学院附属医院产科健康足月分娩新生儿胎盘和脐带血共12份。淋巴细胞亚群检测试剂盒,CD34绝对计数试剂盒(Becton Dickinson公司);CD34磁珠分选试剂盒,FITC标记的CD38单克隆抗体,抗FITC磁珠和MS/LS免疫磁式细胞分选柱(Miltenyi Biotec)。方法:脐带血与RPMI-1640培养基(含体积分数为0.1的胎牛血清)按1∶1的比例混合,采用Ficoll-Histopaque分离液离心30min,吸取界面层细胞,PBS洗涤一次,获得脐带血单个核细胞。采用机械法加0.25g/L胶原酶消化制备胎盘组织单个细胞悬液,之后同脐带血单个核细胞分离步骤分离胎盘单个核细胞。流式细胞仪检测胎盘单个核细胞中CD34+CD38-,CD34+CD38+造血干/祖细胞(HSPCs)和淋巴细胞亚群的组成比例。免疫磁珠分选法分选人胎盘CD34+CD38-、CD34+CD38+造血干/祖细胞,并分别进行粒细胞-单核细胞集落生成单位、红细胞爆裂型集落生成单位、混合集落生成单位系集落形成培养,以评价其造血祖细胞系增殖分化能力。实验全程用脐带血作平行比较分析。主要观察指标:胎盘和脐带血CD34+造血干/祖细胞组成百分率、祖细胞系集落形成能力、淋巴细胞亚群表型及组成特点。结果:①胎盘CD34+造血干/祖细胞百分率是脐带血的8.8倍,差异有显著性BACKGROUND： As is well known that hematopoietic stem and progenitor cells （HSPCs） contain in bone marrow, peripheral blood, and cord blood. Recent studies found that human placenta tissue （PT） also exists in HSPCs. But so far the property and differentiation capacity of human PT-HSPCs is not yet known. Furthermore the composition of lymphocyte subpopulations and immunogenicity regarded to human PT-HSPCs are also unclear. OBJECTIVE： To verify whether there are more HSPCs in human PT than those in human umbilical cord blood （UCB）, to investigate their capacities of proliferation and differentiation, and to analyze the phenotypes of lymphocyte subpopulations in human PT. DESIGN, TIME AND SETTING： Open eXperiments were performed at the Key Laboratory of Cell Engineering of Guizhou Province from January 2004 to December 2006. SETTING： Key Laboratory of Cell Engineering of Guizhou Province, the Affiliated Hospital of Zunyi Medical College. MATERIALS： Twelve human placenta and UCB samples through cesarean delivery were collected aseptically with the informed consents of parturients derived from Maternity Department of the Affiliated Hospital of Zunyi Medical College. The main reagents were detailed as follows： lymphocyte subpopulations analysis reagents Simultest IMK-lymphocyte Kit, CD34 absolute counting reagents Kit （Becton Dickinson）; CD34 Multisort Kit, FITC conjugated CD38 monoclonal antibody, anti-FITC microbeads and MS/LS mini MACS segregating columns （Miltenyi Biotec）. METHODS： UCB samples were 1：1 diluted with RPMI-1640 containing 0.1 volume fraction of fetal bovine serum and the mononuclear cells （MNCs） were isolated on Ficoll-Histopaque by centrifugation for 30 minutes. The MNCs at the interface were collected and washed with PBS. Single cells suspension liquid of human PT was prepared by mechanical method combined with 0.25 g/L collagenase digestion. After that, the placenta samples underwent the same protocol as used in UCB to isolate MNCs. The percentage of CD3

关 键 词：造血干细胞 造血祖细胞 淋巴细胞 集落形成单位 人胎盘 免疫磁式细胞分选 

分 类 号：R394.2[医药卫生—医学遗传学]