六种重组细胞因子对人骨髓造血细胞的体外扩增效应

The Effects of Six Cytokines on the Expansion of Bone Marrow CD34+ Cells in vitro

目的 :观察造血生长因子对骨髓CD34 +细胞的扩增效应及扩增细胞的分化特性。方法 :采用重组人白细胞介素 (rhIL) 1 +rhIL 3+rhIL6 +重组人粒细胞集落刺激因子 +重组人粒 巨噬细胞集落刺激因子和重组人干细胞因子对人骨髓单个核细胞 (MNC)进行刺激 ,动态观察了经上述细胞因子作用后MNC的增殖效应及液态扩增培养后粒 巨噬集落形成率 (CFU GM ) ,利用流式细胞技术 (FACS)分析了扩增前后CD34 +细胞及其亚群的动态变化。结果 :经上述 6种细胞因子作用 2 4d ,骨髓MNC总数是原代MNC的 38.33倍 ,液态扩增 1 0d后 ,CFU GM的数量达高峰 ,是原代MNC的 1 9.0 4倍 ,2 4d时CFU GM总数降至 3.94倍 ,且集落明显小于扩增初期。扩增至 6～ 1 0d ,CD34 +细胞总数是原代MNC的 1 36 .40～ 90 .40倍 ,1 9d时降为 6 1 .40倍。结论 :外源性造血生长因子对CD34 +骨髓细胞具有较强的扩增效应 ,选择适宜的扩增时间是保证骨髓移植后尽快重建造血功能的重要环节。

Purpose:To explore the role of human hematopoietic growth factors (HGFs) in proliferation and differentiation of bone marrow CD34 +cells. Methods: human bone marrow mononuclear cells (MNC) were cultured with 6 cytokines (including rhIL 1, rhIL 3, rhIL 6, rhG CSF, rhGM CSF and rhSCF). The expansion folds of MNC, the CFU GM of hematopoietic cells and the number of CD34 +cells and its subsets were observed. Results: The total nucleated cells were expanded by 38.33 folds after cultured with 6 cytokines for 24 days. The number of CFU GM was increased by 19.04 folds after cultured in liquid for 10 days, and decreased to 3.94 folds after 24 days. The total number of CD34 + cells was increased by 136.40～90.40 folds in 6～10 days, and decreased to 61.40 folds at 19th day. Conclusion: HGFs have significant effects on the expansion of bone marrow CD34 +cells,and culture time is important for the reconstruction of hematopoiesis in bone marrow transplantion.