骨髓基质细胞联合细胞因子对脐血cd133+细胞体外扩增作用的研究.docVIP

骨髓基质细胞联合细胞因子对脐血CD133+细胞体外扩增作用的研究 作者：毛平　曾进龙　王彩霞　杜庆华 【摘要】 　　为了探讨胚胎骨髓基质细胞（FBMSC）联合细胞因子对脐血单个核细胞（MNC）中CD133+细胞的体外扩增作用，将新鲜脐血（CB）中分离出来的MNC接种于无血清培养体系中培养14天。实验分为4组：C组为空白对照组，不含基质细胞和细胞因子；S组为单用基质细胞组；F组为单用细胞因子组；SF组为联合使用基质细胞和细胞因子组。在第0，6，10及14天检测有核细胞总数、CD133+细胞数及集落形成单位（CFU）数。结果表明：各时间点SF组有核细胞总数的扩增倍数均高于其它组；除了第14天外，SF组在第6、10天时CD133+细胞数、CFU数的扩增倍数均高于其它组。结论： 胚胎骨髓基质细胞对延缓造血细胞的分化具有重要的作用，基质细胞联合细胞因子可以有效的扩增脐血单个核细胞及其中的CD133+细胞，这是一种比较接近于临床移植要求的造血细胞体外扩增方法。 【关键词】 骨髓基质细胞； 造血干细胞； 细胞扩增； CD133+细胞； 脐血； 细胞因子 　　In Vitro Expansion of Cord Blood CD133+ Cells Supported by Bone Marrow Stromal Cells and Cytokines Abstract The aim of this study was to investigate the effects of human fetal bone marrow stromal cells (FBMSC) in combination with exogenous cytokines on supporting in vitro expansion of CD133+cells in cord blood mononuclear cells (MNC). MNCs separated from cord blood (CB) were cultured for up to 14 days in a serumfree system with FBMSC or exogenous cytokines or both of them. On day 0,6,10 and 14, total nucleated cells (TNC) were counted；CD133+ cells were quantified by FACS, and hematopoietic progenitor cells were assessed by semisolid culture assay. The results showed that the number of TNC was remarkably increased in FBMSC and cytokine group, the expansion of CD133+ cells and CFU were increased in FBMSC and cytokine group except that on day 14. It is concluded that FBMSC play an important role in delaying the differentiation of hematopoietic cells. FBMSC in combination with exogenous cytokines can promote the effective expansion of CB MNC and CD133+ cells, this expanding system may meet the needs for clinical application of expanded CD133+cells. Key words bone marrow stromal cell; hematopoietic stem cell; cell expansion; CD133+ cell; cord blood; cytokine CD34是目前造血干细胞（HSC）最常用的表面抗原标志。随着对HSC研究的深入，人们发现存在CD34抗原尚未表达的造血干细胞［1］，又发现部分AC133可能是比CD34更早表达的表面抗原［2，3］，在第七届人类白细胞分化抗原大会上AC133正式命名为CD133，CD133+细胞比CD34+细胞具有更高的增殖和自我更新能力［4］。我们利用本实验室已经建立的人胚胎骨髓基质细胞联合细胞因