人胎盘CD133^+、CD133^-细胞组份中高增殖潜能集落形成细胞及其表型特征的初步探讨  

Phenotype property of HPP-CFC in CD133^+ and CD133^- cells subsets derived from human placenta tissues

作　　者：刘佳云[1,2] 陈代雄[1] 章涛[1] 方宁[1] 刘祖林[1] 刘金伟[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,贵州遵义563003 [2]遵义医学院基础医学院病理生理学教研室,贵州遵义563003

出　　处：《遵义医学院学报》2009年第2期118-120,124,共4页Journal of Zunyi Medical University

基　　金：贵州省科技计划发展项目(2051;2003JGY005)

摘　　要：目的研究人胎盘CD133+、CD133-细胞亚群中高增殖潜能集落形成细胞(HPP-CFC)频率及表型特征。方法采用机械法制备人胎盘组织(PT)游离细胞悬液,用Histopaque-1077分离出单个核细胞(MNCs),经磁式分选(MACS)分离纯化CD133+和CD133-细胞。将分选的CD133+和CD133-细胞分别使用H4435培养基培养4wk后观察HPP-CFC集落形成能力,用流式细胞仪(FCM)对分选的细胞组份和HPP-CFC进行表型分析。结果培养4wk后,PT-CD133+、CD133-细胞生成的HPP-CFC集落分别为32.4±11.2/5×103、2.0±2.3/5×103,前者细胞中HPP-CFC数量明显高于后者(P<0.01);PT-CD133+、CD133-细胞培养至28d时CD133+CD34+、CD133+CD34-和CD133-CD34+亚型均比培养前减少。结论人胎盘组织CD133+与CD133-细胞群中均存在HPP-CFC,但CD133+细胞群中的HPP-CFC明显多于CD133-细胞群,说明胎盘CD133+细胞群中存在更多的早期HSPC。Objective To detect frequency of HPP-CFC and analysis the phenotype property of HPP-CFC in CD133^＋ and CD133^-cells derived from human placenta tissues （PT）. Methods Isolated cells suspension liquid of human PT was prepared by mechanical method. Mononucleate eells（MNCs） from PT was separated by Histopaque-1077, then isolating and purifying the different immunophenotype cells subsets CD133^＋ and CD133^- cells contained in MNCs by Magnetic Activated Cell Sorting （MACS）. The CD133^＋ and CD133^- ceils derived from human PT were cultivated for 4 weeks with H4435 medium to investigate capacities of colony forming.The immunophenotype of sorting cells and HPP-CFC were analyzed by flow cytometer （FCM）. Results The number of HPP-CFC in CD133^＋ and CD133^- cells derived from human PT have 32.4± 11.2/5×10^3 and 2.0±2.3/5×10^3 respectively after 4 weeks, The number of HPP-CFC of the former than higher those of the latter obviously （P〈0.01）. It was decreased that the proportion of CD133^＋CD34^＋,CD133^＋CD34^- and CD133^-CD34^＋ cells subsets derived from PT-CD133^＋ and CD133^- cells after culturing 28days compared with 0 day. Conclusions The study presented HPP-CFC both exist in CD133^＋ and CD133^- cells derived from PT.but HPP-CFC exist in PT-CD133^＋ cells than those of CD133^- cells obviously.Which demonstrated more primal HSPCs exist in PT-CD133^＋ cells.

关 键 词：人胎盘 造血干 祖细胞 磁式激活细胞分选 CD133 高增殖潜能集落形成细胞(HPP—CFC) 

分 类 号：Q813.1[生物学—生物工程]