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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。
Objective To investigate the effect of human bone marrow stromal cells (HBMSC) on the expansion of cord blood mononuclear cells (MNCs) in vitro and to find the optimal timing of harvesting. Methods MNC isolated from CB specimens were inoculated into established serum-free culture system (with or without HBMSC and cytokine support). MNC count, number of CD34 + cells and colony forming units (CFU) number. Results In the process of in vitro culture, under the same conditions at all time points, the indexes of HBMSC layer support group were higher than other groups (P <0.05), and HBMSC combined with exogenous cytokine group had the best amplification. In vitro culture 6 ~ 10d, the above indicators basically reached the highest value. Conclusions HBMSC combined with exogenous cytokines can effectively amplify CB in vitro. This system may be an ideal solution to expand hematopoietic stem and progenitor cells. In vitro culture 6 ~ 10d is the best time to harvest.