脐血造血干/祖细胞体外扩增的实验研究

目的探讨因子组合FL+TPO+SCF+IL-6对脐血CD34 +细胞的体外扩增效应.方法采用Mini-MACS分离纯化出CD34+细胞, 在无基质接触的液体悬浮培养体系培养,通过间接免疫荧光染色、流式细胞仪技术、集落形成试验等动态测定细胞扩增情况.结果培养4周后,FL+TPO+SCF+IL-6组细胞总数、集落形成数均比扩增前显著增加(P＜0.05),并且能维持一定比例的CD34+、Th y-1+细胞;SCF+IL-3+IL-6+GM-CSF+EPO组在第2周时集落形成数已降低,第4周时集落形成数、CD34+细胞基本检测不到.结论与经典因子组合SCF+IL-3+IL-6+G M-CSF+EPO相比,因子组合FL+TPO +SCF+IL-6能在较长时间内有效扩增脐血造血干/祖细胞, 是一个优化组合方案.

The Experimental Study on in Vitro Expansion of Human Umbilica l Cord Blood Hematopoietic Stem/Progenitor Cells

Objective To investigate the expansion effects of the cytokine combination (FL TPO SCF IL-6) on purified CD34 cells obtained from cord blood.Methods Cord blood CD34 cells were purified with Mini-MACS and cultured in stroma-free liquid culture system, the expansion effect on CD34 cells was assayed by indirect immunofluorescence staining, flow cytometry, colony-forming test.Results Four weeks later, the number of cells and CFU of group FL TPO SCF IL-6 were increased significantly compared with that in the pre-cultured( P < 0.05 ),there were also some proportional CD34 , Thy-1 cells. The number of CFU reduced in the se cond ,CFC and CD34 cells disappeared in the 4th week.Conclusion The cytokine combination of FL TPO SCF IL-6 can more effectively expand CD34 cells compared with the classical cytokine combination of SCF IL-6 IL-3 GM-CSF EPO and is an optimized project.