脐血造血干/祖细胞体外扩增的实验研究

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

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 .0 5 ) ,并且能维持一定比例的CD34 + 、Thy - 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 +GM -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。

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.