人骨髓基质细胞支持脐血单个核细胞体外扩增

目的探讨人骨髓基质细胞(HBMSC)对脐血(CB)单个核细胞(MNC)体外扩增的支持作用及寻找最佳的收获时机。方法将从CB标本中分离出的MNC接种于已建立的无血清培养体系(含或不含HBMSC及细胞因子支持),在0、6、10及14d检测MNC数、CD34 细胞数及集落形成单位(CFU)数。结果在体外培养过程中,各时间点相同条件下,有HBMSC层支持组的各项测量指标均较其他组高(P<0.05),其中HBMSC联合外源性细胞因子组扩增效果最佳。体外培养6～10d时,以上各项指标基本上达到最高值。结论HBMSC联合外源性细胞因子能实现CB在体外的有效扩增,此体系可能是扩增造血干、祖细胞的较理想方案。体外培养6～10d是收获的最佳时间。

Ex vivo expansion of cord blood mononuclear cells supported by human bone marrow stromal cells

Objective To study the role of human bone marrow stromal cells (HBMSC) in supporting the in vitro expansion of cord blood mononuclear cells (MNCs) and to find the optimal harvest time for clinical use.Methods MNCs separated from cord were cultured in a serum-free system (with HBMSC /cytokines supported or not). On the day 0, 6, 10 and 14, total MNCs were counted, CD34~ cells were quantitated by FACS, and hematopoietic progenitor cells were assessed by semisolid culture assay.Results Compared with other groups, the expansion fold of total cells and CD34~ cells were higher in the group with HBMSC and cytokines supported. After in vitro culture for 6 to 10 days, the above parameters reached the peak values.Conclusions HBMSC in combination with exogenous cytokines could efficiently support hematopoiesis ex vivo for a short-term culture. Days 6~10 may be an optimal harvest time.