骨髓基质细胞联合细胞因子对脐血单个核细胞表面抗原表达的影响

背景：课题组已建立胎儿骨髓基质细胞联合细胞因子的造血细胞体外培养体系,该培养体系能否有效扩增各个发育阶段的造血细胞有待验证。目的：观察骨髓基质细胞联合细胞因子培养体系对脐血单个核细胞表面抗原CD133、CD34表达的影响。方法：将从脐血标本中分离出来的单个核细胞接种于无血清培养体系,实验分为3组：①F组：干细胞因子+Flt3配体+促血小板生成素+单个核细胞。②S组：基质细胞+单个核细胞。③SF组：基质细胞+干细胞因子+Flt3配体+促血小板生成素+单个核细胞。在第0,6,10,14天检测有核细胞总数、CD133⁺、CD34⁺、CD133⁺CD34⁺细胞数以及集落形成单位数。 结果与结论：SF组有核细胞总数在各个检测时间点均比其他两组高;除了第14天外,第6、10天两个时间点SF组中CD133⁺、CD34⁺、CD133⁺CD34⁺细胞及集落形成单位数均高于其他组;含骨髓基质细胞的S组和SF组中CD133⁺细胞/有核细胞、CD34⁺细胞/有核细胞、CD133⁺CD34⁺细胞/有核细胞的比例保持在较高的水平。结果说明骨髓基质细胞联合细胞因子能有效的扩增脐血单个核细胞及其中的CD133⁺、CD34⁺、CD133⁺CD34⁺细胞,基质细胞对维持造血干细胞的原始性具有重要的作用。

Effect of bone marrow stromal cells in combination with exogenous cytokines on antigen expression of cord blood mononuclear cells

BACKGROUND: In vitro hematopoietic culture system of fetal bone marrow stromal cells (BMSCs) combined with exogenous cytokines has been established. It remains to verify whether this culture system can improve the proliferation of hematopoietic cells at various developmental stages.OBJECTIVE:To investigate the effects of the culture system containing BMSCs in combination with exogenous cytokines on umbilical cord blood (UCB) mononuclear cells (MNCs) expressing CD133 and CD34. METHODS:MNCs separated from UCB were cultured in a serum-free system with BMSCs or exogenous cytokines or both of them. On days 0, 6, 10 and 14, total nucleated cells (TNCs) were counted, CD133⁺, CD34⁺, and CD133⁺CD34⁺cells were quantitated by FACS, and hematopoietic progenitor cells were assessed by semisolid culture assay.  RESULTS AND CONCLUSION:The number of TNCs was remarkably more in BMSCs and cytokine group compared with the other two groups. The expression of CD133⁺ and CD133⁺CD34⁺ cells were higher in BMSCs and cytokine group than the other two groups at days 6 and 10. The ratio of CD133⁺, CD34⁺ and CD133⁺CD34⁺ cells was significantly more in the system containing BMSCs. BMSCs in combination with cytokines can expand UCB-MNCs and CD133+, CD34⁺, CD133⁺CD34⁺ cells effectively. BMSCs exert an important role in remaining primitive characteristics of hematooietic stem/progenitor stem cells.