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

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±11．2／5×10^3,2．0±2．3／5×10^3，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。

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

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.