分化抑制培养体系体外扩增脐血造血细胞的研究

Reserch on expansion of cord blood hematopoietic cell by the differentiation inhibitory culture system invitro

目的:探讨分化抑制培养体系对脐血造血细胞体外扩增效应。方法:分化抑制培养体系与脐血单个核细胞(MNC)体外共同培养7 d,检测MNC细胞总数、CFC、CD34+细胞反应扩增效果,并检测CD34+细胞表面归巢相关黏附分子VLA-4(CD49d)、VLA-5(CD49e)、LFA-1(CD11a)、HCAM(CD44)、L-selectin(CD62L)的表达率。结果:分化抑制培养体系组明显扩增脐血MNC细胞总数、CFC、CD34+细胞(均P<0．05),对照组培养脐血MNC细胞总数明显下降,CFC和CD34+细胞完全死亡(均P<0．01)。CD34+细胞表面各黏附分子CD49d、CD44和C1362L表达与扩增前相当(均P>0．05),而CD49e和CD11a表达明显高于扩增前(均P<0．05)。结论:分化抑制培养体系体外显著扩增脐血造血细胞,并且扩增后的造血干(祖)细胞总体上保持其表面归巢相关黏附分子的表达,归巢功能不会减低,是一种安全有效的扩增体系。

Objective：To explore the effect of the differentiation inhibitory culture system on expansion of cord hematopoietic cells ex vivo, and observe the expression of Homing-related cell adhesion molecules VLA4 （CD49d）, VLA-5（CD49e）, LFA-1（CD11a）, HCAM（CD44） and L-selectin（CD62L） on CD34^＋ cells. Method：The differentiation inhibitory culture system was used to culture with cord blood MNC for 7 days, the total number of MNC, CFC CD34^＋ cells were observed, the expression of Homing related Cell adhesion molecules VLA-4 （CD49d）, VLA-5 （CD49e）, LFA-1（CD11a）, HCAM（CD44） and L-selectin（CD62L） on CD34^＋ cells were counted. Result：Tthe differentiation inhibitory culture system significantly enhanced cord blood MNC, CFC, CD34^＋ cells, （ P 〈0.05）. In control group, the total number of MNC was reduced remarkedly, CFC and CD34^＋ cells were completely dead, the majority of cells had apoptosis（ P 〈0.05）. the expression of CD49d, CD44 and CD62L on CD34^＋ cells were similar（ P 〉0.05）, but the expression of CD49e and CDlla on CD34^＋ cells were increased（ P 〈0.05）. Conclusion： the differentiation inhibitory culture system significantly expand cord blood hematopoietic cells Ex Vivo, but the expanded hematopoietic stem/progenitor cell may well sustain the expansion of Homing-related adhesion molecules, homing-function may not reduce, this system is a good expansion system.