脐血造血细胞的体外扩增及分化特征

目的：了解造血生长因子的不同组合对人脐血ＣＤ３４＋细胞的扩增效应及分化特性。方法：采用人脐血单个核细胞（ＭＮＣ）经ｒｈＩＬ－１，ｒｈＩＬ－３，ｒｈＩＬ－６，ｒｈＧ－ＣＳＦ，ｒｈＧＭ－ＣＳＦ和ｒｈＳＣＦ的不同组合进行体外扩增培养，动态观察了不同细胞因子组合对有核细胞总数的扩增数量，应用流式细胞技术（ＦＡＣＳ）动态分析了造血细胞的表面标志，并对液态扩增后造血细胞的粒－巨噬集落形成率（ＣＦＵ－ＧＭ）进行了动态观察。结果：经上述６种细胞因子培养２０ｄ，有核细胞总数增殖４４倍，液态扩增培养８ｄ后，ＣＦＵ－ＧＭ比原代ＭＮＣ增加１４．７４倍，扩增１８ｄ后，ＣＦＵ－ＧＭ明显减少，仅为原代ＭＮＣ的６．４２倍，且集落小于前者。扩增培养至６～８ｄ时，ＣＤ３４＋细胞总数是原代 ＭＮＣ的１２７．７９～１９６．４０倍，１８ｄ时下降至１０１．５１倍。结论：外源性造血生长因子可使脐血ＣＤ３４＋细胞及ＣＦＵ－ＧＭ产生明显的扩增效应。

in vitroExpansion and Differentiation of Cord Blood Hemotopoietic Cells

Objective: To explore the role of human hematopoietic growth factors (HGFs) in proliferation and differentiation of cord blood CD34 cells. Methods: Human cord blood mononuclear cells (MNC) were cultured with different combinations of HGFs (including rhIL-1,rhIL-3, rhIL-6, rhG-CSF, rhGM-CSF and rhSCF). The expansion folds of MNC, the changes of cellular surface markers by FACS, and CFU-GM of hematopoietic cells were observed. Results: The total nucleated cells expanded by 44 folds after cultured with 6 cytokines for 20 days. The number of CFU-GM increased by 14 .74 folds after cultured in liquid for 8 days, but decreased heavily after 18 days. The total number of CD34 cells increased by 127. 79 ~ 196 .40 folds at 6 to 8 day, but decreased to 101.51 folds at 18 day. Conclusion: Human hematopoietic growth factors can increase the expansion of cord blood CD34 cells and CFU-GM significantly.