脐血造血细胞体外集落培养最佳收获时机的探讨

目的 探讨脐血（CB）多系造血集落培养在不同培养条件下脐血造血细胞体外扩增的最佳收获时机。方法 将从新鲜CB标本中分离出的单个核细胞（MNC）分别接种于已建立的无血清培养基，该培养基分别含单用人骨髓基质细胞、单用细胞因子及细胞因子联合入骨髓基质细胞（HBMSC）支持培养体系。在0、6、10及14d各时间点分别取细胞进行巨细胞集落形成单位（CFU—MK）、红乐爆式集落形成单位（BFU—E）及粒单细胞集落形成单位（CFU—GM）培养。结果在体外培养过程中，第10d时各组CFU—MK、BFU—E及CFU—GM数均高于组内其它时间点（P〈0．05）；在含HBMSC支持培养的组集落生成数优于其它组（P〈0．05）。结论 脐血MNC体外培养过程中，第10d可能是收获的最佳时机。FIBMSC能更好地维持造血干／祖细胞的活性，使其集落形成能数得到有效扩增。

Observation on the optimum time for harvesting hematopoietic cells from cord blood after culture in vitro

Objective To explore the optimal time for harvesting cord blood hematopoietic cells cultured in vitro by colony multiplication. Methods Mononuclear cells separated from cord blood were cultured in a serum-free system supplemented with cytokines or human bone marrow stromal cells of combination of them. CFU-MK,BFU-E and CFU-Gm were assessed by semisolid culture assay on d0,d6,d10 and d14 for determination of progenitor unmber. Results The quantity of CFU-MK,BFU-E and CFU-Gm on d10 after culture were significantly higher than that of other culture time (P<0.05). The number of colony forming units were higher in the groups supported with human bone marrow stromal cells (P<0.05). Conclusion The optimal time for harvesting the mononuclear cells is on the tenth day after in vitro culture of cord blood hematopoietic cells. Bone marrow stromal cells is capable of maintaining the activity of hematopoietic cells cultured in vitro.