人脐血CD133细胞的分离及其体外扩增的研究  

Study on Separation of CD133 Cells From Human Cord Blood and Their Amplification in Vitro

作　　者：谭玲[1] 陈勇华[1] 张春明[1] 陈伟[1] 沈亚芳[1] 沈鹤柏[1] 董亚明[1] 李兴玉[1] 

机构地区：[1]上海师范大学生命与环境科学学院化学系,上海200234

出　　处：《现代生物医学进展》2007年第6期853-856,共4页Progress in Modern Biomedicine

摘　　要：目的：探讨免疫磁性纳米粒子分离人脐血CD133细胞的方法,了解分离出的CD133细胞在体外短期培养中的变化及其在体外扩增的可能性.方法：通过化学沉淀法制备具有超顺磁性的r-Fe2O3纳米粒子,在其表面包裹具有生物亲合性的二氧化硅,并在其表面通过化学修饰使其成为生物功能化的磁性纳米粒子.再通过一定的化学连接方法将单克隆抗体CD133连接到生物功能化的磁性纳米粒子表面使其成为免疫磁性纳米粒子,然后利用自制的免疫磁性纳米粒子从单个核细胞中分离出CD133细胞,并分别对单个核细胞和CD133细胞在体外短期培养中的动态变化进行了初步观察和比较.结果：经免疫磁性纳米粒子分离的脐血中CD133细胞平均数为（5±1.4）×107/ml,占单个核细胞数的（3±0.3）%;单个核细胞（对照组）和CD133细胞（实验组）分别进行红、粒系集落扩增培养14天、21天,实验组中两种造血祖细胞集落扩增倍数都明显高于对照组（P＜0.01）.结论：使用自制的免疫磁性纳米粒子能较好的分离脐血中的CD133细胞,分离与纯化出来的CD133细胞不仅细胞活力不受影响,而且与单个核细胞相比具有更强的增殖能力.Objective： To separate the CD133 cells from human cord blood （HCB） by using immunomagnetic nanoparfides and to investigate the amplification of CD133 cells form HCB in vitro. Methods： The superparamagnefic nanopaticles were synthesized by the classic coprecipitaton method .And the biological compatble silica was coated to the surface of these magnetic nanoparticles .The biofunctonal magnetic nanoparticles were obtained by modifying the nanoparticles with the amino group. The antibody was combined to the nanoparticles by chemical method. The immunomagnetic nanoparficles were applied in the separation Of CD133 cell from the Cord Blood. Results： The mean value of CD133 cells in cord blood separated by immunomagnetic nanoparticles was （5＋1.4） ×107/ml, which accounted for （3±0.3）%of the number of mononuclear cells （MNC）. MNC （control group） and CD133 cells （experimental group） were respectively cultured in CFU-E and CFU-GM for 14d and 21d, the amplification of CFU-E and CFU-GM in experimental group was markedly higher than those in control group （P〈0.01）. Conclusions： The immunomagnetc nanoparficles were able to separate the CD133 cells from the human Cord Blood .For the separated and purified cells,their activity was not affected and had stronger proliferation compared with MNC.

关 键 词：CD133 造血干细胞 免疫磁性纳米粒子 

分 类 号：R392.1[医药卫生—免疫学]